Total synthesis of trioxacarcin dc-45-a2 and preparation of trioxacarcin analogs

ABSTRACT

In one aspect, the present invention provides novel derivatives of trioxacarin analogs of the formula (I) wherein the variables are as defined herein. The application also provides compositions, methods of treatment, and methods of synthesis thereof.

This application claims the benefit of priority to U.S. ProvisionalApplication Ser. No. 62/094,662, filed on Dec. 19, 2014 and U.S.Provisional Application Ser. No. 62/186,128, filed Jun. 29, 2015, theentire contents of which are hereby incorporated by reference.

BACKGROUND 1. Field

This disclosure relates to the fields of medicine, pharmacology,chemistry, antimicrobial activity, and oncology. In particular, newcompounds, compositions, methods of treatment, and methods of synthesisrelating to trioxacarcin and derivatives thereof are disclosed.

2. Related Art

A group of natural products and synthetic derivatives thereof known asantitumor antibiotics are known to be powerful chemotherapeutics. Thesecompounds have a variety of different mechanisms for their cytotoxicity,but often are associated with modification to the cellular DNA. Severalcommercial chemotherapeutics including actinomycin, bleomycin,daunorubicin, mitoxantrone, and doxorubicin fall within this class ofcompounds. One particular natural product, trioxacarcin DC-45-A2 (1,FIG. 1), is a naturally occurring antitumor antibiotic that serves as abiosynthetic precursor to a variety of other biologically active membersof the family, including the highly potent DC-45-A1 (2), trioxacarcin A(3), and LL-D49194α1 (4) (FIG. 1) (Tomita et al., 1981; Tamaoki et al.,1981; Maiese et al., 1990; Maskey et al., 2004; Shirahata et al., 1984).The complex architecture with multiple oxygen containing functionalgroups and numerous stereocenters presents a difficult syntheticchallenge limiting its commercial viability (Cassidy et al., 1993; Sunet al., 1994; Smith et al., 1995; Maskey et al., 2004; Fitzner et al.,2008; Pfoh et al., 2008). While originally these compounds were obtainedby fermentation, several different synthetic routes have been developed(Gaoni, 1968; Waserman et al., 1969; Wasserman et al., 1986a; Wassermanet al., 1986b; Wasserman et al., 1988a; Wasserman et al., 1988b; Naruseet al., 1988a; Naruse et al., 1988b; Evans et al., 1991). Unfortunately,these methods are still relatively difficult and require numerousdifferent steps to obtain the desired final product without allowingaccess to other derivatives. As such, analogs of trioxacarcin as well asan improved and modular synthesis method which allows for easier accessto the natural product and analogs thereof are of commercial interest.

SUMMARY

Thus, there is provided compounds of the formula:

wherein: R₁ is amino, hydroxy, or mercapto; alkoxy_((C≤12)),cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),acyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), acylthio_((C≤12)),alkylamino_((C≤12)), cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),alkynylamino_((C≤12)), dialkylamino_((C≤12)),dicycloalkylamino_((C≤12)), dialkenylamino_((C≤12)),dialkynylamino_((C≤12)), amido_((C≤12)), or a substituted version of anyof these groups; or R₁ is a group of the formula:—O-alkanediyl_((C≤8))-alkoxy_((C≤12)),—O-alkanediyl_((C≤8))-alkenyloxy_((C≤12)),—O-alkanediyl_((C≤8))-alkynyloxy_((C≤12)), or a substituted versionthereof; or R₁ is a group of the formula:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein: R_(a) is hydrogen, alkyl_((C≤6)), or substituted alkyl_((C≤6));R₂ and R₃ are independently hydrogen, amino, hydroxy, mercapto;alkyl_((C≤12)), cycloalkyl_((C≤12)), alkenyl_((C≤12)), alkynyl_((C≤12)),alkoxy_((C≤12)), cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)),alkynyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), alkylamino_((C≤12)),cycloalkylamino_((C≤12)), alkenylamino_((C≤12)), alkynylamino_((C≤12)),or a substituted version of any of these groups; R₂ and R₃ are takentogether and are alkoxydiyl_((C≤8)), alkylaminodiyl_((C≤12)),alkylthiodiyl_((C≤12)), or a substituted version of any of these groups;R₄ is hydrogen, amino, halo, hydroxy, mercapto, alkyl_((C≤12)) orsubstituted alkyl_((C≤12)); X₁ and X₂ are each independently hydrogen,hydroxy, or alkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),or a substituted version of any of these groups; and A is a fusedcycloalkanediyl and has the structure:

wherein: Y₁ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₁ is oxo, then the atom to which Y₁is bound is part of a double bond, and provided that when the atom towhich Y₁ is bound is part of a double bond, then Y₁ is oxo; Y₂ ishydrogen, hydroxy, alkyl_((C≤12)), substituted alkyl_((C≤12)),alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or —OX₃, wherein X₃ is ahydroxy protecting group; or a group of the formula:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein: R_(a) is hydrogen, alkyl_((C≤6)), or substituted alkyl_((C≤6));and n₁ is 0, 1, 2, 3, 4, 5, or 6; or A is a fused arenediyl and has thestructure:

wherein: Y₃ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₄ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; and n₂ is 0, 1, 2,or 3; or A is a fused arenediyl with a fused heterocycloalkanediyl andhas the structure:

wherein: Y₅ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₆ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; Y₇ is hydrogen,alkyl_((C≤12)), or substituted alkyl_((C≤12)); n₃ is 0 or 1; and x is 1,2, 3, or 4; or A is a fused heteroarenediyl and has the structure:

wherein: Z₁, Z₂, and Z₃ are each independently selected from CR₅R₅′,NR₅″, O, or S; R₅ and R₅′ are each independently hydrogen, amino,hydroxy, halo, cyano, nitro, sulfato, sulfamido; alkyl_((C≤6)),alkoxy_((C≤6)), alkylamino_((C≤6)), dialkylamino_((C≤12)),amido_((C≤6)), or a substituted version of any of these groups; and R₅″is hydrogen, alkyl_((C≤12)), or substituted alkyl_((C≤12)); providedthat at least one of Z₁, Z₂, or Z₃ is NR₅″, O, or S; n₄ is 1, 2, 3, or4; or A is a fused arenediyl with a fused cycloalkanediyl and has thestructure:

wherein: Y₈ and Y₉ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₀ is hydrogen, oxo, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; n₅ is 0 or 1; and y is 0, 1, 2, 3, 4, 5, 6, 7, or 8; A is a fusedarenediyl and has the structure:

wherein: Y₁₁ and Y₁₂ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₃ is hydrogen, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; and n₆ is 0, 1, 2, 3, or 4; provided that R₁ is not hydroxy andeither R₂ or R₃ is methoxy when A is a fused cycloalkanediyl of theformula:

or a pharmaceutically acceptable salt thereof. In some embodiments, thecompounds are further defined as:

wherein: R₁ is amino, hydroxy, or mercapto; alkoxy_((C≤12)),cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),acyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), acylthio_((C≤12)),alkylamino_((C≤12)), cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),alkynylamino_((C≤12)), dialkylamino_((C≤12)),dicycloalkylamino_((C≤12)), dialkenylamino_((C≤12)),dialkynylamino_((C≤12)), amido_((C≤12)), or a substituted version of anyof these groups; or R₁ is a group of the formula:—O-alkanediyl_((C≤8))-alkoxy_((C≤12)),—O-alkanediyl_((C≤8))-alkenyloxy_((C≤12)),—O-alkanediyl_((C≤8))-alkynyloxy_((C≤12)), or a substituted versionthereof; or R₁ is a group of the formula:

wherein: R₆, R₇, R₈, and R₉ are each independently hydrogen, hydroxy,alkyl_((C≤8)), alkoxy_((C≤8)), substituted alkyl_((C≤8)), or substitutedalkoxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; R₂and R₃ are independently selected from hydrogen, amino, hydroxy,mercapto; alkyl_((C≤12)), cycloalkyl_((C≤12)), alkenyl_((C≤12)),alkynyl_((C≤12)), alkoxy_((C≤12)), cycloalkoxy_((C≤12)),alkenyloxy_((C≤12)), alkynyloxy_((C≤12)), alkylthio_((C≤12)),cycloalkylthio_((C≤12)), alkenylthio_((C≤12)), alkynylthio_((C≤12)),alkylamino_((C≤12)), cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),alkynylamino_((C≤12)), or a substituted version of any of these groups;R₂ and R₃ are taken together and are alkoxydiyl_((C≤8)),alkylaminodiyl_((C≤12)), alkylthiodiyl_((C≤12)), or a substitutedversion of any of these groups; R₄ is hydrogen, amino, halo, hydroxy,mercapto, alkyl_((C≤12)) or substituted alkyl_((C≤12)); X₁ and X₂ areeach independently hydrogen, hydroxy, or alkoxy_((C≤12)),alkenyloxy_((C≤12)), alkynyloxy_((C≤12)), or a substituted version ofany of these groups; and A is a fused cycloalkanediyl and has thestructure:

wherein: Y₁ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₁ is oxo, then the atom to which Y₁is bound is part of a double bond, and provided that when the atom towhich Y₁ is bound is part of a double bond, then Y₁ is oxo; Y₂ ishydrogen, hydroxy, alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or—OX₃, wherein X₃ is a hydroxy protecting group; and n₁ is 0, 1, 2, 3, 4,5, or 6; or A is a fused arenediyl and has the structure:

wherein: Y₃ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₄ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; and n₂ is 0, 1, 2,or 3; or A is a fused arenediyl with a fused heterocycloalkanediyl andhas the structure:

wherein: Y₅ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₆ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; Y₇ is hydrogen,alkyl_((C≤12)), or substituted alkyl_((C≤12)); n₃ is 0 or 1; and x is 1,2, 3, or 4; or A is a fused heteroarenediyl and has the structure:

wherein: Z₁, Z₂, and Z₃ are each independently selected from CR₅R₅′,NR₅″, O, or S; R₅ and R₅′ are each independently hydrogen, amino,hydroxy, halo, cyano, nitro, sulfato, sulfamido; alkyl_((C≤6)),alkoxy_((C≤6)), alkylamino_((C≤6)), dialkylamino_((C≤12)),amido_((C≤6)), or a substituted version of any of these groups; and R₅″is hydrogen, alkyl_((C≤12)), or substituted alkyl_((C≤12)); providedthat at least one of Z₁, Z₂, or Z₃ is NR₅″, O, or S; n₄ is 1, 2, 3, or4; or A is a fused arenediyl with a fused cycloalkanediyl and has thestructure:

wherein: Y₈ and Y₉ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₀ is hydrogen, oxo, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; n₅ is 0 or 1; and y is 0, 1, 2, 3, 4, 5, 6, 7, or 8; A is a fusedarenediyl and has the structure:

wherein: Y₁₁ and Y₁₂ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₃ is hydrogen, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; and n₆ is 0, 1, 2, 3, or 4; provided that R₁ is not hydroxy andeither R₂ or R₃ is methoxy when A is a fused cycloalkanediyl of theformula:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the formula is further defined as Ia. In someembodiments, the formula is further defined as Ib. In some embodiments,the formula is further defined as Ic. In some embodiments, the formulais further defined as Id. In some embodiments, the formula is furtherdefined as Ie. In some embodiments, the formula is further defined asIf.

The compound according to any one of claims 1-7, wherein R₁ is:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein: R_(a) is hydrogen, alkyl_((C≤6)), or substituted alkyl_((C≤6)).In some embodiments, R₁ is:

wherein: R₆, R₇, R₈, and R₉ are each independently hydrogen, hydroxy,alkyl_((C≤8)), alkoxy_((C≤8)), substituted alkyl_((C≤8)), or substitutedalkoxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group. Insome embodiments, the thiol reactive group of R₁₂ is a maleimide. Inother embodiments, R₁ is a group of the formula:—O-alkanediyl_((C≤8))-alkoxy_((C≤12)),—O-alkanediyl_((C≤8))-alkenyloxy_((C≤12)),—O-alkanediyl_((C≤8))-alkynyloxy_((C≤12)), or a substituted versionthereof. In some embodiments, the alkanediyl_((C≤8)) of R₁ is —CH₂—. Inother embodiments, R₁ is alkoxy_((C≤12)) or substituted alkoxy_((C≤12)).In some embodiments, R₁ is alkoxy_((C≤12)). In some embodiments, R₁ issubstituted alkoxy_((C≤12)). In some embodiments, R₁ is —O(CH₂)₆OH or—OCH₂CH₂SH. In some embodiments, R₁ is alkynyloxy_((C≤12)) orsubstituted alkynyloxy_((C≤12)). In some embodiments, R₁ isalkynyloxy_((C≤12)). In some embodiments, R₁ is —CH₂CCH. In someembodiments, R₁ is alkylthio_((C≤12)) or substituted alkylthio_((C≤12)).In some embodiments, R₁ is alkylthio_((C≤12)). In some embodiments, R₁is —SCH₂CH₃.

In some embodiments, R₂ is hydrogen. In some embodiments, R₂ isalkyl_((C≤12)) or substituted alkyl_((C≤12)). In some embodiments, R₂ isalkyl_((C≤12)). In some embodiments, R₂ is methyl. In some embodiments,R₂ is alkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In someembodiments, R₂ is alkoxy_((C≤12)). In some embodiments, R₂ is methoxy.In some embodiments, R₂ is alkylthio_((C≤12)) or substitutedalkylthio_((C≤12)). In some embodiments, R₂ is alkylthio_((C≤12)). Insome embodiments, R₂ is —SCH₃.

In some embodiments, R₂ and R₃ are taken together and isalkoxydiyl_((C≤12)) or substituted alkoxydiyl_((C≤12)). In someembodiments, R₂ and R₃ are taken together and are alkoxydiyl_((C≤12)).In some embodiments, R₂ and R₃ are —OCH₂CH₂O—, —OCH₂CH₂CH₂O—, or—OCH₂C(CH₃)₂CH₂O—. In other embodiments, R₂ and R₃ are taken togetherand are alkoxydiyl_((C≤12)). In some embodiments, R₂ and R₃ are—OCH₂CH(CH₂OH)CH₂O—, —OCH₂CH(CH₂SH)CH₂O—, or —OCH₂CH(CH₂NHAc)CH₂O—. Inother embodiments, R₂ and R₃ are taken together and isalkylthiodiyl_((C≤12)) or substituted alkylthiodiyl_((C≤12)). In someembodiments, R₂ and R₃ are taken together and arealkylthiodiyl_((C≤12)). In some embodiments, R₂ and R₃ are —SCH₂CH₂CH₂S—or —SCH₂C(CH₃)₂CH₂S—.

In some embodiments, R₃ is hydrogen. In some embodiments, R₃ isalkyl_((C≤12)) or substituted alkyl_((C≤12)). In some embodiments, R₃ isalkyl_((C≤12)). In some embodiments, R₃ is methyl. In some embodiments,R₃ is alkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In someembodiments, R₃ is alkoxy_((C≤12)). In some embodiments, R₃ is methoxy.In some embodiments, R₃ is alkylthio_((C≤12)) or substitutedalkylthio_((C≤12)). In some embodiments, R₃ is alkylthio_((C≤12)). Insome embodiments, R₃ is —SCH₃.

In some embodiments, R₄ is halo. In some embodiments, R₄ is fluoro,chloro, or bromo. In some embodiments, R₄ is fluoro. In someembodiments, R₄ is alkyl_((C≤12)) or substituted alkyl_((C≤12)). In someembodiments, R₄ is alkyl_((C≤12)). In some embodiments, R₄ is methyl. Insome embodiments, R₄ is substituted alkyl_((C≤12)). In some embodiments,R₄ is trifluoromethyl.

In some embodiments, X₁ is hydrogen. In some embodiments, X₁ is hydroxy.In some embodiments, X₁ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)). In some embodiments, X₁ is alkoxy_((C≤12)). In someembodiments, X₁ is methoxy. In other embodiments, X₁ is substitutedalkoxy_((C≤12)). In some embodiments, X₁ is —O(CH₂)₃NH₂,—O(CH₂)₂C(O)NH₂, or —O(CH₂)₃SH. In other embodiments, X₁ isalkenyloxy_((C≤12)). In some embodiments, X₁ is —OCH₂CHCH₂. In otherembodiments, X₁ is alkynyloxy_((C≤12)). In some embodiments, X₁ is—OCH₂CCH.

In some embodiments, X₂ is hydrogen. In some embodiments, X₂ is hydroxy.In some embodiments, X₂ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)). In some embodiments, X₂ is alkoxy_((C≤12)). In someembodiments, X₂ is methoxy.

In some embodiments, Y₁ is oxo. In some embodiments, Y₂ is hydrogen. Insome embodiments, Y₂ is hydroxy. In some embodiments, 76.1. The compoundaccording to any one of claim 1-2 or 12-74, wherein Y₂ is:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein: R_(a) is hydrogen, alkyl_((C≤6)), or substituted alkyl_((C≤6)).In some embodiments, Y₂ is:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyloxy_((C≤12)), acyl_((C≤12)), substitutedalkyl_((C≤12)), substituted alkoxy_((C≤12)), substitutedacyloxy_((C≤12)), or substituted acyl_((C≤12)). In some embodiments, Y₂is:

In some embodiments, n₁ is 0, 1, 2, or 3. In some embodiments, n₁ is 0,1, or 2.

In some embodiments, Y₃ is hydroxy. In some embodiments, Y₃ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In some embodiments, Y₃is alkoxy_((C≤12)). In some embodiments, Y₃ is methoxy. In someembodiments, Y₃ is substituted alkoxy_((C≤12)). In some embodiments, Y₃is methoxymethoxy. In some embodiments, Y₄ is hydrogen. In someembodiments, Y₄ is hydroxy. In some embodiments, Y₄ is alkoxy_((C≤12))or substituted alkoxy_((C≤12)). In some embodiments, Y₄ isalkoxy_((C≤12)). In some embodiments, Y₄ is methoxy. In someembodiments, Y₄ is alkylamino_((C≤12)) or substitutedalkylamino_((C≤12)). In some embodiments, Y₄ is alkylamino_((C≤12)). Insome embodiments, Y₄ is methylamino. In some embodiments, n₂ is 1, 2, or3.

In some embodiments, Y₅ is hydroxy. In some embodiments, Y₅ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In some embodiments, Y₅is alkoxy_((C≤12)). In some embodiments, Y₅ is methoxy. In someembodiments, Y₆ is hydrogen. In some embodiments, Y₆ is hydroxy. In someembodiments, Y₆ is alkoxy_((C≤12)) or substituted alkoxy_((C≤12)). Insome embodiments, Y₆ is alkoxy_((C≤12)). In some embodiments, Y₆ ismethoxy. In some embodiments, Y₇ is hydrogen. In some embodiments, Y₇ isalkyl_((C≤6)) or substituted alkyl_((C≤6)). In some embodiments, x is 2or 3. In some embodiments, x is 2. In some embodiments, x is 3. In someembodiments, n₃ is 0. In some embodiments, n₃ is 1.

In some embodiments, Z₁ is S. In some embodiments, Z₁ is N. In someembodiments, Z₁ is O. In some embodiments, Z₂ is S. In some embodiments,Z₂ is N. In some embodiments, Z₂ is O. In some embodiments, Z₂ is CR₅″.In some embodiments, R₅″ is hydrogen, hydroxy, halo, alkyl_((C≤12)),substituted alkyl_((C≤12)), alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)). In some embodiments, R₅″ is hydrogen. In someembodiments, R₅″ is alkoxy_((C≤12)) or substituted alkoxy_((C≤12)). Insome embodiments, R₅″ is methoxy. In some embodiments, R₅″ isalkyl_((C≤12)) or substituted alkyl_((C≤12)). In some embodiments, R₅″is methyl. In some embodiments, Z₃ is S. In some embodiments, Z₃ is N.In some embodiments, Z₃ is O. In some embodiments, Z₃ is CR₅″. In someembodiments, R₅″ is hydrogen, hydroxy, halo, alkyl_((C≤12)), substitutedalkyl_((C≤12)), alkoxy_((C≤12)), or substituted alkoxy_((C≤12)). In someembodiments, R₅″ is hydrogen. In some embodiments, R₅″ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In some embodiments, R₅″is methoxy. In some embodiments, R₅″ is alkyl_((C≤12)) or substitutedalkyl_((C≤12)). In some embodiments, R₅″ is methyl. In some embodiments,n₄ is 1, 2, or 3.

In some embodiments, Y₈ is hydrogen. In some embodiments, Y₈ is hydroxy.In some embodiments, Y₈ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)). In some embodiments, Y₈ is alkoxy_((C≤12)). In someembodiments, Y₈ is methoxy. In some embodiments, Y₉ is hydrogen. In someembodiments, Y₉ is hydroxy. In some embodiments, Y₉ is alkoxy_((C≤12))or substituted alkoxy_((C≤12)). In some embodiments, Y₉ isalkoxy_((C≤12)). In some embodiments, Y₉ is methoxy. In someembodiments, Y₁₀ is hydrogen. In some embodiments, Y₁₀ is hydroxy. Insome embodiments, Y₁₀ is oxo. In some embodiments, Y₁₀ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In some embodiments, Y₁₀is alkoxy_((C≤12)). In some embodiments, Y₁₀ is methoxy. In someembodiments, Y₁₀ is alkylamino_((C≤12)) or substitutedalkylamino_((C≤12)). In some embodiments, Y₁₀ is alkylamino_((C≤12)). Insome embodiments, Y₁₀ is methylamino. In some embodiments, n₅ is 1. Insome embodiments, y is 1, 2, 3, 4, 5, or 6.

In some embodiments, Y₁₁ is hydrogen. In some embodiments, Y₁₁ ishydroxy. In some embodiments, Y₁₁ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)). In some embodiments, Y₁₁ is alkoxy_((C≤12)). In someembodiments, Y₁₁ is methoxy. In some embodiments, Y₁₂ is hydrogen. Insome embodiments, Y₁₂ is hydroxy. In some embodiments, Y₁₂ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In some embodiments, Y₁₂is alkoxy_((C≤12)). In some embodiments, Y₁₂ is methoxy. In someembodiments, Y₁₃ is hydrogen. In some embodiments, Y₁₃ is hydroxy. Insome embodiments, Y₁₃ is oxo. In some embodiments, Y₁₃ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)). In some embodiments, Y₁₃is alkoxy_((C≤12)). In some embodiments, Y₁₃ is methoxy. In someembodiments, Y₁₃ is alkylamino_((C≤12)) or substitutedalkylamino_((C≤12)). In some embodiments, Y₁₃ is alkylamino_((C≤12)). Insome embodiments, Y₁₃ is methylamino. In some embodiments, n₆ is 1, 2,or 3. In some embodiments, n₆ is 2 or 3.

In some embodiments, the compound is further defined as:

or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present disclosure provides pharmaceuticalcompositions comprising a compound of the present disclosure and apharmaceutically acceptable carrier. In some embodiments, thecomposition is formulated for administration: orally, intraadiposally,intraarterially, intraarticularly, intracranially, intradermally,intralesionally, intramuscularly, intranasally, intraocularly,intrapericardially, intraperitoneally, intrapleurally,intraprostatically, intrarectally, intrathecally, intratracheally,intratumorally, intraumbilically, intravaginally, intravenously,intravesicularlly, intravitreally, liposomally, locally, mucosally,parenterally, rectally, subconjunctival, subcutaneously, sublingually,topically, transbuccally, transdermally, vaginally, in crémes, in lipidcompositions, via a catheter, via a lavage, via continuous infusion, viainfusion, via inhalation, via injection, via local delivery, or vialocalized perfusion.

In yet another aspect, the present disclosure provides methods oftreating a disease or disorder in a patient in need thereof comprisingadministering to the patient a pharmaceutically effective amount of acompound or composition of the present disclosure. In some embodiments,the disease or disorder is cancer. In some embodiments, the cancer is acarcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiplemyeloma, or seminoma. In some embodiments, the cancer is of the bladder,blood, bone, brain, breast, central nervous system, cervix, colon,endometrium, esophagus, gall bladder, gastrointestinal tract, genitalia,genitourinary tract, head, kidney, larynx, liver, lung, muscle tissue,neck, oral or nasal mucosa, ovary, pancreas, prostate, skin, spleen,small intestine, large intestine, stomach, testicle, or thyroid. In someembodiments, the method further comprises a second therapeutic agent ormodality. In some embodiments, the compound is administered once. Insome embodiments, the compound is administered two or more times.

In yet another aspect, the present disclosure provides methods oftreating a disease or disorder in a patient in need thereof comprisingadministering to the patient a pharmaceutically effective amount of acompound or composition of the present disclosure. In some embodiments,the disease or disorder is a bacterial infection, a parasitic infection,or a viral infection. In some embodiments, the disease or disorder is abacteria infection wherein the bacteria is a gram positive bacteria. Inother embodiments, the disease or disorder is a bacteria infectionwherein the bacteria is a gram negative bacteria. In some embodiments,the disease is a parasitic infection. In some embodiments, the parasiticinfection causes malaria.

In still another aspect, the present disclosure provides methods ofpreparing a compound comprising reacting a compound of the formula:

wherein: R₁ is amino, hydroxy, or mercapto; or —OX₃, wherein X₃ is ahydroxy protecting group, —SX₄, wherein X₄ is a thio protecting group,or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protectinggroup and the other is a hydrogen or X₅ and X₆ are taken together andare a divalent amine protecting group; R₂ and R₃ are independentlyselected from hydrogen, amino, hydroxy, mercapto; alkyl_((C≤12)),cycloalkyl_((C≤12)), alkenyl_((C≤12)), alkynyl_((C≤12)),alkoxy_((C≤12)), cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)),alkynyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), alkylamino_((C≤12)),cycloalkylamino_((C≤12)), alkenylamino_((C≤12)), alkynylamino_((C≤12)),or a substituted version of any of these groups, or —OX₇, wherein X₇ isa hydroxy protecting group, —SX₈, wherein X₈ is a thio protecting group,or —NX₉X₁₀, wherein either X₉ or X₁₀ is a monovalent amine protectinggroup and the other is a hydrogen or X₉ and X₁₀ are taken together andare a divalent amine protecting group; R₂ and R₃ are taken together andare alkoxydiyl_((C≤8)), alkylaminodiyl_((C≤12)), alkylthiodiyl_((C≤12)),or a substituted version of any of these groups; R₄ is hydrogen, amino,halo, hydroxy, mercapto, alkyl_((C≤12)) or substituted alkyl_((C≤12)),or —OX₁₁, wherein X₁₁ is a hydroxy protecting group, —SX₁₂, wherein X₁₂is a thio protecting group, or —NX₁₃X₁₄, wherein either X₁₃ or X₁₄ is amonovalent amine protecting group and the other is a hydrogen or X₁₃ andX₁₄ are taken together and are a divalent amine protecting group; R₅ ishydrogen or a hydroxy protecting group; R₆ is hydrogen oralkylidene_((C≤12)), alkyl_((C≤12)), cycloalkyl_((C≤12)),alkenyl_((C≤12)), alkynyl_((C≤12)), alkoxy_((C≤12)),cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),alkylamino_((C≤12)), dialkylamino_((C≤12)), or a substituted version ofany of these groups X₁ and X₂ are each independently hydrogen, hydroxy,alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or —OX₁₅, wherein X₁₅ is ahydroxy protecting group; or X₁₅ and R₅ are taken together and are adivalent diol protecting group; and A is a fused cycloalkanediyl and hasthe structure:

wherein: Y₁ is hydrogen, oxo, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), or —OX₁₆, wherein X₁₆ is a hydroxy protecting group;provided that when Y₁ is oxo, then the atom to which Y₁ is bound is partof a double bond, and provided that when the atom to which Y₁ is boundis part of a double bond, then Y₁ is oxo; Y₂ is hydrogen, hydroxy,alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or —OX₁₇, wherein X₁₇ is ahydroxy protecting group; and n₁ is 0, 1, 2, 3, 4, 5, or 6; or A is afused arenediyl and has the structure:

wherein: Y₃ is hydrogen, oxo, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), or —OX₁₈, wherein X₁₈ is a hydroxy protecting group;provided that when Y₃ is oxo, then the atom to which Y₃ is bound is partof a double bond, and provided that when the atom to which Y₃ is boundis part of a double bond, then Y₃ is oxo; Y₄ is hydrogen, hydroxy,amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₁₉, wherein X₁₉ is a hydroxyprotecting group, —SX₂₀, wherein X₂₀ is a thio protecting group, or—NX₂₁X₂₂, wherein either X₂₁ or X₂₂ is a monovalent amine protectinggroup and the other is a hydrogen or X₂₁ and X₂₂ are taken together andare a divalent amine protecting group; and n₂ is 0, 1, 2, or 3; or A isa fused arenediyl with a fused heterocycloalkanediyl and has thestructure:

wherein: Y₅ is hydrogen, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),or —OX₂₃, wherein X₂₃ is a hydroxy protecting group; Y₆ is hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₂₄, wherein X₂₄ is a hydroxyprotecting group, —SX₂₅, wherein X₂₅ is a thio protecting group, or—NX₂₆X₂₇, wherein either X₂₆ or X₂₇ is a monovalent amine protectinggroup and the other is a hydrogen or X₂₆ and X₂₇ are taken together andare a divalent amine protecting group; Y₇ is hydrogen, alkyl_((C≤12)),or substituted alkyl_((C≤12)); n₃ is 0 or 1; and x is 1, 2, 3, or 4; orA is a fused heteroarenediyl and has the structure:

wherein: Z₁, Z₂, and Z₃ are each independently selected from CR₇R₇′,NR₈, O, or S; R₇ and R₇′ are each independently hydrogen, amino,hydroxy, halo, cyano, nitro, sulfato, sulfamido; alkyl_((C≤6)),alkoxy_((C≤6)), alkylamino_((C≤6)), dialkylamino_((C≤12)),amido_((C≤6)), or a substituted version of any of these groups; and R₈is hydrogen, alkyl_((C≤12)), or substituted alkyl_((C≤12)); providedthat at least one of Z₁, Z₂, or Z₃ is NR₇, O, or S; n₄ is 1, 2, 3, or 4;or A is a fused arenediyl with a fused cycloalkanediyl and has thestructure:

wherein: Y₈ and Y₉ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₂₈, wherein X₂₈ is a hydroxyprotecting group, —SX₂₉, wherein X₂₉ is a thio protecting group, or—NX₃₀X₃₁, wherein either X₃₀ or X₃₁ is a monovalent amine protectinggroup and the other is a hydrogen or X₃₀ and X₃₁ are taken together andare a divalent amine protecting group; Y₁₀ is hydrogen, oxo, hydroxy,amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃₂, wherein X₃₂ is a hydroxyprotecting group, —SX₃₃, wherein X₃₃ is a thio protecting group, or—NX₃₄X₃₅, wherein either X₃₄ or X₃₅ is a monovalent amine protectinggroup and the other is a hydrogen or X₃₄ and X₃₅ are taken together andare a divalent amine protecting group, provided that when Y₁₀ is oxo,then the atom to which Y₁₀ is bound is part of a double bond, andprovided that when the atom to which Y₁₀ is bound is part of a doublebond, then Y₁₀ is oxo; n₅ is 0 or 1; and y is 0, 1, 2, 3, 4, 5, 6, 7, or8; A is a fused arenediyl and has the structure:

wherein: Y₁₁ and Y₁₂ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃₆, wherein X₃₆ is a hydroxyprotecting group, —SX₃₇, wherein X₃₇ is a thio protecting group, or—NX₃₈X₃₉, wherein either X₃₈ or X₃₉ is a monovalent amine protectinggroup and the other is a hydrogen or X₃₈ and X₃₉ are taken together andare a divalent amine protecting group; Y₁₃ is hydrogen, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₄₀, wherein X₄₀ is a hydroxyprotecting group, —SX₄₁, wherein X₄₁ is a thio protecting group, or—NX₄₂X₄₃, wherein either X₄₂ or X₄₃ is a monovalent amine protectinggroup and the other is a hydrogen or X₄₂ and X₄₃ are taken together andare a divalent amine protecting group; and n₆ is 0, 1, 2, 3, or 4; witha Lewis acid under conditions sufficient to produce a compound of theformula:

wherein: R₁, R₂, R₃, R₄, R₅, R₆, X₁, and X₂ are as defined above; A is afused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

orwherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; or a salt thereof.In some embodiments, the Lewis acid is a transition metal or a boroncomplex. In some embodiments, the Lewis acid is a boron complex. In someembodiments, the Lewis acid is boron trifluoride etherate. In someembodiments, the Lewis acid is a transition metal complex. In someembodiments, the Lewis acid is SnCl₄. In some embodiments, the methodfurther comprises a solvent. In some embodiments, the solvent ischloroalkane_((C≤12)). In some embodiments, the solvent isdichloromethane. In some embodiments, R₆ is alkylidene_((C≤12)) orsubstituted alkyldiene_((C≤12)). In some embodiments, the method furthercomprises reacting the compound with an epoxidizing agent underconditions to sufficient to produce a compound of the formula:

wherein: R₁, R₂, R₃, R₄, R₅, X₁, and X₂ are as defined above; A is afused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

orwherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; In someembodiments, the epoxidizing agent is osmium tetraoxide with tosylchloride and a base. In some embodiments, the osmium tetraoxide is addedto the compound and after a time period of about 1 hour to about 24hours, the tosyl chloride and the base are added. In some embodiments,the method further comprises reacting the compound with an oxidizingagent under conditions sufficient to produce a compound of the formula:

wherein: R₂, R₃, R₄, R₅, X₁, and X₂ are as defined above; R₁ is O, S, orNH; and A is a fused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

orwherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; or a salt thereof.In some embodiments, the oxidizing agent is tetrapropylammoniumperruthenate and N-methylmorpholine N-oxide.

In some embodiments, the method further comprises reacting the compoundwith a fluoride source under condition sufficient to produce a compoundof the formula:

wherein: R₂, R₃, R₄, X₁, and X₂ are as defined above; R₁ is amino,hydroxy, or mercapto; alkoxy_((C≤12)), cycloalkoxy_((C≤12)),alkenyloxy_((C≤12)), alkynyloxy_((C≤12)), alkylthio_((C≤12)),cycloalkylthio_((C≤12)), alkenylthio_((C≤12)), alkynylthio_((C≤12)),alkylamino_((C≤12)), cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),alkynylamino_((C≤12)), dialkylamino_((C≤12)),dicycloalkylamino_((C≤12)), dialkenylamino_((C≤12)),dialkynylamino_((C≤12)), or a substituted version of any of thesegroups; A is a fused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

wherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; or a salt thereof.In some embodiments, the fluoride source is Et₃N.3HF. In someembodiments, the method further comprises one or more deprotectionsteps.

In another aspect, the present disclosure provides conjugates of theformula:

(A-L)_(n)-X  (VI)

wherein: A is a compound described herein; L is a covalent bond or alinker; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and X is a celltargeting moiety.

In some embodiments, one or more steps of the reaction further comprisespurifying the reaction in a purification step. In some embodiments, thepurification method is chromatography. In some embodiments, thepurification method is column chromatography or high performance liquidchromatography.

It is contemplated that any method or composition described herein canbe implemented with respect to any other method or composition describedherein. For example, an aldehyde synthesized by one method may be usedin the preparation of a final compound according to a different method.

The use of the word “a” or “an” when used in conjunction with the term“comprising” in the claims and/or the specification may mean “one,” butit is also consistent with the meaning of “one or more,” “at least one,”and “one or more than one.” The word “about” means plus or minus 5% ofthe stated number.

Other objects, features and advantages of the present disclosure willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and the specificexamples, while indicating specific embodiments of the disclosure, aregiven by way of illustration only, since various changes andmodifications within the spirit and scope of the disclosure will becomeapparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE FIGURES

The following drawings form part of the present specification and areincluded to further demonstrate certain aspects of the presentinvention. The invention may be better understood by reference to one ormore of these drawings in combination with the detailed description.

FIG. 1—Molecular structures of trioxacarcins: DC-45-A2 (1), DC-45-A1(2), A (3) and LL-D49194α1 (4).

FIG. 2—Retrosynthetic analysis of trioxacarcin DC-45-A2 (1).TBS=tert-butyldimethylsilyl, PMB=p-methoxybenzyl, TMS=trimethylsilyl,MOM=methoxymethyl.

FIGS. 3A-3C—Graph of luminescence as a function of compoundconcentration for Trox4-Trox9 in cytotoxity assay for (FIG. 3A) MES SA,(FIG. 3B) MES SA DX, and (FIG. 3C) 293T.

FIGS. 4A-4C—Graph of luminescence as a function of compoundconcentration for Trox12-Trox16 in cytotoxity assay for (FIG. 4A) MESSA, (FIG. 4B) MES SA DX, and (FIG. 4C) 293T.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The present disclosure relates to a modular synthesis of trioxacarcinDC-45-A2 and development of trioxacarcin analogs. In some aspects, thepresent disclosure provides novel analogs of trioxacarcin which may beuseful in the treatment of proliferative diseases such as cancer.Without wishing to be bound by any theory, trioxacarcin are antitumorantibiotics. In yet another aspect, the present disclosure relates to amodular synthesis which incorporates a macrocyclic rearrangement whichconstructs the dioxabicyclo[2.2.1]heptane. These and other aspects ofthe disclosure are described in greater detail below.

I. COMPOUNDS AND FORMULATIONS THEREOF

A. Compounds

In one aspect, the present disclosure provides compounds of the formula:

wherein:

-   -   R₁ is amino, hydroxy, or mercapto;    -   alkoxy_((C≤12)), cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)),        alkynyloxy_((C≤12)), acyloxy_((C≤12)), alkylthio_((C≤12)),        cycloalkylthio_((C≤12)), alkenylthio_((C≤12)),        alkynylthio_((C≤12)), acylthio_((C≤12)), alkylamino_((C≤12)),        cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),        alkynylamino_((C≤12)), dialkylamino_((C≤12)),        dicycloalkylamino_((C≤12)), dialkenylamino_((C≤12)),        dialkynylamino_((C≤12)), amido_((C≤12)), or a substituted        version of any of these groups; or    -   R₁ is a group of the formula:        —O-alkanediyl_((C≤8))-alkoxy_((C≤12)),        —O-alkanediyl_((C≤8))-alkenyloxy_((C≤12)),        —O-alkanediyl_((C≤8))-alkynyloxy_((C≤12)), or a substituted        version thereof; or    -   R₁ is a group of the formula:

-   -   wherein:        -   R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independently            hydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)),            acyloxy_((C≤8)), substituted alkyl_((C≤8)), substituted            alkoxy_((C≤8)), or substituted acyloxy_((C≤8)); and        -   R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)), alkoxy_((C≤12)),            acyl_((C≤12)), substituted alkyl_((C≤12)), substituted            alkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of            the formula:

-   -   -   wherein:            -   R₁₁ is hydrogen, alkyl_((C≤8)), or substituted                alkyl_((C≤8)); and            -   R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substituted                alkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive                group, or a substituted version of                —O-alkanediyl_((C≤12))-a thiol reactive group; or        -   R₇ and R₁₀ are taken together to form a heterocyclic            compound of the formula:

-   -   -   wherein:            -   R_(a) is hydrogen, alkyl_((C≤6)), or substituted                alkyl_((C≤6));

    -   R₂ and R₃ are independently hydrogen, amino, hydroxy, mercapto;

    -   alkyl_((C≤12)), cycloalkyl_((C≤12)), alkenyl_((C≤12)),        alkynyl_((C≤12)), alkoxy_((C≤12)), cycloalkoxy_((C≤12)),        alkenyloxy_((C≤12)), alkynyloxy_((C≤12)), alkylthio_((C≤12)),        cycloalkylthio_((C≤12)), alkenylthio_((C≤12)),        alkynylthio_((C≤12)), alkylamino_((C≤12)),        cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),        alkynylamino_((C≤12)), or a substituted version of any of these        groups;

    -   R₂ and R₃ are taken together and are alkoxydiyl_((C≤8)),        alkylaminodiyl_((C≤12)), alkylthiodiyl_((C≤12)), or a        substituted version of any of these groups;

    -   R₄ is hydrogen, amino, halo, hydroxy, mercapto, alkyl_((C≤12))        or substituted alkyl_((C≤12));

    -   X₁ and X₂ are each independently hydrogen, hydroxy, or        -   alkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),            or a substituted version of any of these groups; and

    -   A is a fused cycloalkanediyl and has the structure:

-   -   wherein:        -   Y₁ is hydrogen, oxo, alkoxy_((C≤12)), or substituted            alkoxy_((C≤12)), provided that when Y₁ is oxo, then the atom            to which Y₁ is bound is part of a double bond, and provided            that when the atom to which Y₁ is bound is part of a double            bond, then Y₁ is oxo;        -   Y₂ is hydrogen, hydroxy, alkyl_((C≤12)), substituted            alkyl_((C≤12)), alkoxy_((C≤12)), substituted            alkoxy_((C≤12)), or —OX₃, wherein X₃ is a hydroxy protecting            group; or a group of the formula:

-   -   -   wherein:            -   R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independently                hydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)),                acyloxy_((C≤8)), substituted alkyl_((C≤8)), substituted                alkoxy_((C≤8)), or substituted acyloxy_((C≤8)); and            -   R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),                alkoxy_((C≤12)), acyl_((C≤12)), substituted                alkyl_((C≤12)), substituted alkoxy_((C≤12)), substituted                acyl_((C≤12)), or a group of the formula:

-   -   -   -   wherein:                -   R₁₁ is hydrogen, alkyl_((C≤8)), or substituted                    alkyl_((C≤8)); and                -   R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)),                    substituted alkoxy_((C≤12)), —O—                    alkanediyl_((C≤12))-a thiol reactive group, or a                    substituted version of —O-alkanediyl_((C≤12))-a                    thiol reactive group; or            -   R₇ and R₁₀ are taken together to form a heterocyclic                compound of the formula:

-   -   -   -   wherein:                -   R_(a) is hydrogen, alkyl_((C≤6)), or substituted                    alkyl_((C≤6)); and

        -   n₁ is 0, 1, 2, 3, 4, 5, or 6; or            A is a fused arenediyl and has the structure:

-   -   wherein:        -   Y₃ is hydrogen, oxo, alkoxy_((C≤12)), or substituted            alkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom            to which Y₃ is bound is part of a double bond, and provided            that when the atom to which Y₃ is bound is part of a double            bond, then Y₃ is oxo;        -   Y₄ is hydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)),            substituted alkoxy_((C≤12)), alkylthio_((C≤12)), substituted            alkylthio_((C≤12)), alkylamino_((C≤12)), substituted            alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxy            protecting group, —SX₄, wherein X₄ is a thio protecting            group, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent            amine protecting group and the other is a hydrogen or X₅ and            X₆ are taken together and are a divalent amine protecting            group; and        -   n₂ is 0, 1, 2, or 3; or            A is a fused arenediyl with a fused heterocycloalkanediyl            and has the structure:

-   -   wherein:        -   Y₅ is hydrogen, oxo, alkoxy_((C≤12)), or substituted            alkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom            to which Y₃ is bound is part of a double bond, and provided            that when the atom to which Y₃ is bound is part of a double            bond, then Y₃ is oxo;        -   Y₆ is hydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)),            substituted alkoxy_((C≤12)), alkylthio_((C≤12)), substituted            alkylthio_((C≤12)), alkylamino_((C≤12)), substituted            alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxy            protecting group, —SX₄, wherein X₄ is a thio protecting            group, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent            amine protecting group and the other is a hydrogen or X₅ and            X₆ are taken together and are a divalent amine protecting            group;        -   Y₇ is hydrogen, alkyl_((C≤12)), or substituted            alkyl_((C≤12));        -   n₃ is 0 or 1; and        -   x is 1, 2, 3, or 4; or            A is a fused heteroarenediyl and has the structure:

-   -   wherein:        -   Z₁, Z₂, and Z₃ are each independently selected from CR₅R₅′,            NR₅″, O, or S;            -   R₅ and R₅′ are each independently hydrogen, amino,                hydroxy, halo, cyano, nitro, sulfato, sulfamido;                -   alkyl_((C≤6)), alkoxy_((C≤6)), alkylamino_((C≤6)),                    dialkylamino_((C≤12)), amido_((C≤6)), or a                    substituted version of any of these groups; and            -   R₅″ is hydrogen, alkyl_((C≤12)), or substituted                alkyl_((C≤12));            -   provided that at least one of Z₁, Z₂, or Z₃ is NR₅″, O,                or S;        -   n₄ is 1, 2, 3, or 4; or    -   A is a fused arenediyl with a fused cycloalkanediyl and has the        structure:

-   -   wherein:        -   Y₈ and Y₉ are each independently selected from hydrogen,            hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted            alkoxy_((C≤12)), alkylthio_((C≤12)), substituted            alkylthio_((C≤12)), alkylamino_((C≤12)), substituted            alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxy            protecting group, —SX₄, wherein X₄ is a thio protecting            group, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent            amine protecting group and the other is a hydrogen or X₅ and            X₆ are taken together and are a divalent amine protecting            group;        -   Y₁₀ is hydrogen, oxo, hydroxy, amino, mercapto,            alkoxy_((C≤12)), substituted alkoxy_((C≤12)),            alkylthio_((C≤12)), substituted alkylthio_((C≤12)),            alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃,            wherein X₃ is a hydroxy protecting group, —SX₄, wherein X₄            is a thio protecting group, or —NX₅X₆, wherein either X₅ or            X₆ is a monovalent amine protecting group and the other is a            hydrogen or X₅ and X₆ are taken together and are a divalent            amine protecting group, provided that when Y₃ is oxo, then            the atom to which Y₃ is bound is part of a double bond, and            provided that when the atom to which Y₃ is bound is part of            a double bond, then Y₃ is OXO;        -   n₅ is 0 or 1; and        -   y is 0, 1, 2, 3, 4, 5, 6, 7, or 8;            A is a fused arenediyl and has the structure:

-   -   wherein:        -   Y₁₁ and Y₁₂ are each independently selected from hydrogen,            hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted            alkoxy_((C≤12)), alkylthio_((C≤12)), substituted            alkylthio_((C≤12)), alkylamino_((C≤12)), substituted            alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxy            protecting group, —SX₄, wherein X₄ is a thio protecting            group, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent            amine protecting group and the other is a hydrogen or X₅ and            X₆ are taken together and are a divalent amine protecting            group;        -   Y₁₃ is hydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)),            substituted alkoxy_((C≤12)), alkylthio_((C≤12)), substituted            alkylthio_((C≤12)), alkylamino_((C≤12)), substituted            alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxy            protecting group, —SX₄, wherein X₄ is a thio protecting            group, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent            amine protecting group and the other is a hydrogen or X₅ and            X₆ are taken together and are a divalent amine protecting            group, provided that when Y₃ is oxo, then the atom to which            Y₃ is bound is part of a double bond, and provided that when            the atom to which Y₃ is bound is part of a double bond, then            Y₃ is oxo; and        -   n₆ is 0, 1, 2, 3, or 4;    -   provided that R₁ is not hydroxy and either R₂ or R₃ is methoxy        when A is a fused cycloalkanediyl of the formula:

or a pharmaceutically acceptable salt thereof.

Additionally, the compounds provided by the present disclosure areshown, for example, above in the summary of the invention section and inthe examples and claims below. They may be made using the methodsoutlined in the Examples section. Trioxacarcin and derivatives thereofcan be synthesized according to the methods described, for example, inthe Examples section below. These methods can be further modified andoptimized using the principles and techniques of organic chemistry asapplied by a person skilled in the art. Such principles and techniquesare taught, for example, in March's Advanced Organic Chemistry:Reactions, Mechanisms, and Structure (2007), which is incorporated byreference herein.

Trioxacarcin and derivatives of the disclosure may contain one or moreasymmetrically-substituted carbon or nitrogen atoms, and may be isolatedin optically active or racemic form. Thus, all chiral, diastereomeric,racemic form, epimeric form, and all geometric isomeric forms of achemical formula are intended, unless the specific stereochemistry orisomeric form is specifically indicated. Compounds may occur asracemates and racemic mixtures, single enantiomers, diastereomericmixtures and individual diastereomers. In some embodiments, a singlediastereomer is obtained. The chiral centers of the compounds of thepresent invention can have the S or the R configuration.

Chemical formulas used to represent trioxacarcin and derivatives thereofof the present disclosure will typically only show one of possiblyseveral different tautomers. For example, many types of ketone groupsare known to exist in equilibrium with corresponding enol groups.Similarly, many types of imine groups exist in equilibrium with enaminegroups. Regardless of which tautomer is depicted for a given compound,and regardless of which one is most prevalent, all tautomers of a givenchemical formula are intended.

Trioxacarcin and derivatives thereof of the present disclosure may alsohave the advantage that they may be more efficacious than, be less toxicthan, be longer acting than, be more potent than, produce fewer sideeffects than, be more easily absorbed than, and/or have a betterpharmacokinetic profile (e.g., higher oral bioavailability and/or lowerclearance) than, and/or have other useful pharmacological, physical, orchemical properties over, compounds known in the prior art, whether foruse in the indications stated herein or otherwise.

In addition, atoms making up trioxacarcin and derivatives thereof of thepresent disclosure are intended to include all isotopic forms of suchatoms. Isotopes, as used herein, include those atoms having the sameatomic number but different mass numbers. By way of general example andwithout limitation, isotopes of hydrogen include tritium and deuterium,and isotopes of carbon include ¹³C and ¹⁴C.

Trioxacarcin and derivatives thereof of the present disclosure may alsoexist in prodrug form. Since prodrugs are known to enhance numerousdesirable qualities of pharmaceuticals (e.g., solubility,bioavailability, manufacturing, etc.), the compounds employed in somemethods of the disclosure may, if desired, be delivered in prodrug form.Thus, the invention contemplates prodrugs of compounds of the presentinvention as well as methods of delivering prodrugs. Prodrugs oftrioxacarcin and derivatives thereof employed in the disclosure may beprepared by modifying functional groups present in the compound in sucha way that the modifications are cleaved, either in routine manipulationor in vivo, to the parent compound. Accordingly, prodrugs include, forexample, compounds described herein in which a hydroxy, amino, orcarboxy group is bonded to any group that, when the prodrug isadministered to a subject, cleaves to form a hydroxy, amino, orcarboxylic acid, respectively.

It should be recognized that the particular anion or cation forming apart of any salt form of a compound provided herein is not critical, solong as the salt, as a whole, is pharmacologically acceptable.Additional examples of pharmaceutically acceptable salts and theirmethods of preparation and use are presented in Handbook ofPharmaceutical Salts: Properties, and Use (2002), which is incorporatedherein by reference.

Those skilled in the art of organic chemistry will appreciate that manyorganic compounds can form complexes with solvents in which they arereacted or from which they are precipitated or crystallized. Thesecomplexes are known as “solvates.” For example, a complex with water isknown as a “hydrate.” Solvates of trioxacarcin and its derivativesprovided herein are within the scope of the invention. It will also beappreciated by those skilled in organic chemistry that many organiccompounds can exist in more than one crystalline form. For example,crystalline form may vary from solvate to solvate. Thus, all crystallineforms of trioxacarcin and derivatives thereof or the pharmaceuticallyacceptable solvates thereof are within the scope of the presentinvention.

B. Formulations

In some embodiments of the present disclosure, the compounds areincluded a pharmaceutical formulation. Materials for use in thepreparation of microspheres and/or microcapsules are, e.g.,biodegradable/bioerodible polymers such as polygalactin, poly-(isobutylcyanoacrylate), poly(2-hydroxyethyl-L-glutamine) and, poly(lactic acid).Biocompatible carriers that may be used when formulating a controlledrelease parenteral formulation are carbohydrates (e.g., dextrans),proteins (e.g., albumin), lipoproteins, or antibodies. Materials for usein implants can be non-biodegradable (e.g., polydimethyl siloxane) orbiodegradable (e.g., poly(caprolactone), poly(lactic acid),poly(glycolic acid) or poly(ortho esters) or combinations thereof).

Formulations for oral use include tablets containing the activeingredient(s) (e.g., trioxacarcin and its derivatives) in a mixture withnon-toxic pharmaceutically acceptable excipients. Such formulations areknown to the skilled artisan. Excipients may be, for example, inertdiluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol,microcrystalline cellulose, starches including potato starch, calciumcarbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate,or sodium phosphate); granulating and disintegrating agents (e.g.,cellulose derivatives including microcrystalline cellulose, starchesincluding potato starch, croscarmellose sodium, alginates, or alginicacid); binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginicacid, sodium alginate, gelatin, starch, pregelatinized starch,microcrystalline cellulose, magnesium aluminum silicate,carboxymethylcellulose sodium, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethyleneglycol); and lubricating agents, glidants, and antiadhesives (e.g.,magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenatedvegetable oils, or talc). Other pharmaceutically acceptable excipientscan be colorants, flavoring agents, plasticizers, humectants, bufferingagents, and the like.

The tablets may be uncoated or they may be coated by known techniques,optionally to delay disintegration and absorption in thegastrointestinal tract and thereby providing a sustained action over alonger period. The coating may be adapted to release the active drug ina predetermined pattern (e.g., in order to achieve a controlled releaseformulation) or it may be adapted not to release the active drug untilafter passage of the stomach (enteric coating). The coating may be asugar coating, a film coating (e.g., based on hydroxypropylmethylcellulose, methylcellulose, methyl hydroxyethylcellulose,hydroxypropylcellulose, carboxymethylcellulose, acrylate copolymers,polyethylene glycols and/or polyvinylpyrrolidone), or an enteric coating(e.g., based on methacrylic acid copolymer, cellulose acetate phthalate,hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcelluloseacetate succinate, polyvinyl acetate phthalate, shellac, and/orethylcellulose). Furthermore, a time delay material, such as, e.g.,glyceryl monostearate or glyceryl distearate may be employed.

II. BACTERIAL INFECTIONS

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection. While humans containnumerous different bacteria on and inside their bodies, an imbalance inbacterial levels or the introduction of pathogenic bacteria can cause asymptomatic bacterial infection. Pathogenic bacteria cause a variety ofdifferent diseases including but not limited to numerous foodborneillness, typhoid fever, tuberculosis, pneumonia, syphilis, and leprosy.

Additionally, different bacteria have a wide range of interactions withbody and those interactions can modulate ability of the bacteria tocause an infection. For example, bacteria can be conditionallypathogenic such that they only cause an infection under specificconditions. For example, Staphylococcus and Streptococcus bacteria existin the normal human bacterial biome, but these bacteria when they areallowed to colonize other parts of the body causing a skin infection,pneumonia, or sepsis. Other bacteria are known as opportunisticpathogens and only cause diseases in a patient with a weakened immunesystem or another disease or disorder.

Bacteria can also be intracellular pathogens which can grow andreproduce within the cells of the host organism. Such bacteria can bedivided into two major categories as either obligate intracellularparasites or facultative intracellular parasites. Obligate intracellularparasites require the host cell in order to reproduce and include suchbacteria as but are not limited to Chlamydophila, Rickettsia, andEhrlichia which are known to cause pneumonia, urinary tract infections,typhus, and Rocky Mountain spotted fever. Facultative intracellularparasites can reproduce either intracellular or extracellular. Somenon-limiting examples of facultative intracellular parasites includeSalmonella, Listeria, Legionella, Mycobacterium, and Brucella which areknown to cause food poisoning, typhoid fever, sepsis, meningitis,Legionnaire's disease, tuberculosis, leprosy, and brucellosis.

Finally, bacterial infections could be targeted to a specific locationin or on the body. For example, bacteria could be harmless if onlyexposed to the specific organs, but when it comes in contact with aspecific organ or tissue, the bacteria can begin replicating and cause abacterial infection.

A. Gram-Positive Bacteria

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection by a gram-positivebacteria. Gram-positive bacteria contain a thick peptidoglycan layerwithin the cell wall which prevents the bacteria from releasing thestain when dyed with crystal violet. Without being bound by theory, thegram-positive bacteria are often more susceptible to antibiotics.Generally, gram-positive bacteria, in addition to the thickpeptidoglycan layer, also comprise a lipid monolayer and containteichoic acids which react with lipids to form lipoteichoic acids thatcan act as a chelating agent. Additionally, in gram-positive bacteria,the peptidoglycan layer is outer surface of the bacteria. Manygram-positive bacteria have been known to cause disease including, butare not limited to, Streptococcus, Straphylococcus, Corynebacterium,Enterococcus, Listeria, Bacillus, Clostridium, Rathybacter, Leifsonia,and Clavibacter.

B. Gram-Negative Bacteria

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection by a gram-negativebacteria. Gram-negative bacteria do not retain the crystal violet stainafter washing with alcohol. Gram-negative bacteria, on the other hand,have a thin peptidoglycan layer with an outer membrane oflipopolysaccharides and phospholipids as well as a space between thepeptidoglycan and the outer cell membrane called the periplasmic space.Gram-negative bacterial generally do not have teichoic acids orlipoteichoic acids in their outer coating. Generally, gram-negativebacteria also release some endotoxin and contain prions which act asmolecular transport units for specific compounds. Most bacteria aregram-negative. Some non-limiting examples of gram-negative bacteriainclude Bordetella, Borrelia, Burcelia, Campylobacteria, Escherichia,Francisella, Haemophilus, Helicobacter, Legionella, Leptospira,Neisseria, Pseudomonas, Rickettsia, Salmonella, Shigella, Treponema,Vibrio, and Yersinia.

C. Gram-Indeterminate Bacteria

In some aspects of the present disclosure, the compounds disclosedherein may be used to treat a bacterial infection by agram-indeterminate bacteria. Gram-indeterminate bacteria do not fullstain or partially stain when exposed to crystal violet. Without beingbound by theory, a gram-indeterminate bacteria may exhibit some of theproperties of the gram-positive and gram-negative bacteria. Anon-limiting example of a gram-indeterminate bacteria includemycobacterium tuberculosis or mycobacterium leprae.

III. HYPERPROLIFERATIVE DISEASES

A. Cancer and Other Hyperproliferative Disease

While hyperproliferative diseases can be associated with any diseasewhich causes a cell to begin to reproduce uncontrollably, theprototypical example is cancer. One of the key elements of cancer isthat the cell's normal apoptotic cycle is interrupted and thus agentsthat interrupt the growth of the cells are important as therapeuticagents for treating these diseases. In this disclosure, the trioxacarcinand derivatives thereof may be used to lead to decreased cell counts andas such can potentially be used to treat a variety of types of cancerlines. In some aspects, it is anticipated that the trioxacarcin andderivatives thereof of the present disclosure may be used to treatvirtually any malignancy.

Cancer cells that may be treated with the compounds of the presentdisclosure include but are not limited to cells from the bladder, blood,bone, bone marrow, brain, breast, colon, esophagus, gastrointestine,gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate,skin, stomach, pancreas, testis, tongue, cervix, or uterus. In addition,the cancer may specifically be of the following histological type,though it is not limited to these: neoplasm, malignant; carcinoma;carcinoma, undifferentiated; giant and spindle cell carcinoma; smallcell carcinoma; papillary carcinoma; squamous cell carcinoma;lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma;transitional cell carcinoma; papillary transitional cell carcinoma;adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma;hepatocellular carcinoma; combined hepatocellular carcinoma andcholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma;adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposiscoli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolaradenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma;acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clearcell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma;papillary and follicular adenocarcinoma; nonencapsulating sclerosingcarcinoma; adrenal cortical carcinoma; endometroid carcinoma; skinappendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma;ceruminous adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma;papillary cystadenocarcinoma; papillary serous cystadenocarcinoma;mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cellcarcinoma; infiltrating duct carcinoma; medullary carcinoma; lobularcarcinoma; inflammatory carcinoma; Paget's disease, mammary; acinar cellcarcinoma; adenosquamous carcinoma; adenocarcinoma w/squamousmetaplasia; thymoma, malignant; ovarian stromal tumor, malignant;thecoma, malignant; granulosa cell tumor, malignant; androblastoma,malignant; sertoli cell carcinoma; leydig cell tumor, malignant; lipidcell tumor, malignant; paraganglioma, malignant; extra-mammaryparaganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignantmelanoma; amelanotic melanoma; superficial spreading melanoma; malignantmelanoma in giant pigmented nevus; epithelioid cell melanoma; bluenevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma,malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma;embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma;mixed tumor, malignant; Mullerian mixed tumor; nephroblastoma;hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; Brenner tumor,malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma,malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant;struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant;hemangiosarcoma; hemangioendothelioma, malignant; Kaposi's sarcoma;hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma;juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant;mesenchymal chondrosarcoma; giant cell tumor of bone; Ewing's sarcoma;odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma,malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma;glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma;fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma;oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma;ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactoryneurogenic tumor; meningioma, malignant; neurofibrosarcoma;neurilemmoma, malignant; granular cell tumor, malignant; malignantlymphoma; Hodgkin's disease; paragranuloma; malignant lymphoma, smalllymphocytic; malignant lymphoma, large cell, diffuse; malignantlymphoma, follicular; mycosis fungoides; other specified non-Hodgkin'slymphomas; malignant histiocytosis; multiple myeloma; mast cell sarcoma;immunoproliferative small intestinal disease; leukemia; lymphoidleukemia; plasma cell leukemia; erythroleukemia; lymphosarcoma cellleukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia;monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia;myeloid sarcoma; and hairy cell leukemia. In certain aspects, the tumormay comprise an osteosarcoma, angiosarcoma, rhabdosarcoma,leiomyosarcoma, Ewing sarcoma, glioblastoma, neuroblastoma, or leukemia.

IV. CELL TARGETING MOIETIES

In some aspects, the present disclosure provides compounds conjugateddirectly or through linkers to a cell targeting moiety. In someembodiments, the conjugation of the compound to a cell targeting moietyincreases the efficacy of the compound in treating a disease ordisorder. Cell targeting moieties according to the embodiments may be,for example, an antibody, a growth factor, a hormone, a peptide, anaptamer, a small molecule such as a hormone, an imaging agent, orcofactor, or a cytokine. For instance, a cell targeting moiety accordingthe embodiments may bind to a liver cancer cell such as a Hep3B cell. Ithas been demonstrated that the gp240 antigen is expressed in a varietyof melanomas but not in normal tissues. Thus, in some embodiments, thecompounds of the present disclosure may be used in conjugates with anantibody for a specific antigen that is expressed by a cancer cell butnot in normal tissues.

In certain additional embodiments, it is envisioned that cancer celltargeting moieties bind to multiple types of cancer cells. For example,the 8H9 monoclonal antibody and the single chain antibodies derivedtherefrom bind to a glycoprotein that is expressed on breast cancers,sarcomas and neuroblastomas (Onda et al., 2004). Another example is thecell targeting agents described in U.S. Patent Publication No.2004/005647 and in Winthrop et al. (2003) that bind to MUC-1, an antigenthat is expressed on a variety cancer types. Thus, it will be understoodthat in certain embodiments, cell targeting constructs according theembodiments may be targeted against a plurality of cancer or tumortypes.

Additionally, certain cell surface molecules are highly expressed intumor cells, including hormone receptors such as human chorionicgonadotropin receptor and gonadotropin releasing hormone receptor(Nechushtan et al., 1997). Therefore, the corresponding hormones may beused as the cell-specific targeting moieties in cancer therapy.Additionally, the cell targeting moiety that may be used include acofactor, a sugar, a drug molecule, an imaging agent, or a fluorescentdye. Many cancerous cells are known to over express folate receptors andthus folic acid or other folate derivatives may be used as conjugates totrigger cell-specific interaction between the conjugates of the presentdisclosure and a cell (Campbell, et al., 1991; Weitman, et al., 1992).

Since a large number of cell surface receptors have been identified inhematopoietic cells of various lineages, ligands or antibodies specificfor these receptors may be used as cell-specific targeting moieties.IL-2 may also be used as a cell-specific targeting moiety in a chimericprotein to target IL-2R+ cells. Alternatively, other molecules such asB7-1, B7-2 and CD40 may be used to specifically target activated T cells(The Leucocyte Antigen Facts Book, 1993, Barclay et al. (eds.), AcademicPress). Furthermore, B cells express CD19, CD40 and IL-4 receptor andmay be targeted by moieties that bind these receptors, such as CD40ligand, IL-4, IL-5, IL-6 and CD28. The elimination of immune cells suchas T cells and B cells is particularly useful in the treatment oflymphoid tumors.

Other cytokines that may be used to target specific cell subsets includethe interleukins (IL-1 through IL-15), granulocyte-colony stimulatingfactor, macrophage-colony stimulating factor, granulocyte-macrophagecolony stimulating factor, leukemia inhibitory factor, tumor necrosisfactor, transforming growth factor, epidermal growth factor,insulin-like growth factors, and/or fibroblast growth factor (Thompson(ed.), 1994, The Cytokine Handbook, Academic Press, San Diego). In someaspects, the targeting polypeptide is a cytokine that bind to the Fn14receptor, such as TWEAK (see, e.g., Winkles, 2008; Zhou et al., 2011 andBurkly et al., 2007, incorporated herein by reference).

A skilled artisan recognizes that there are a variety of knowncytokines, including hematopoietins (four-helix bundles) (such as EPO(erythropoietin), IL-2 (T-cell growth factor), IL-3 (multicolony CSF),IL-4 (BCGF-1, BSF-1), IL-5 (BCGF-2), IL-6 IL-4 (IFN-β2, BSF-2, BCDF),IL-7, IL-8, IL-9, IL-11, IL-13 (P600), G-CSF, IL-15 (T-cell growthfactor), GM-CSF (granulocyte macrophage colony stimulating factor), OSM(OM, oncostatin M), and LIF (leukemia inhibitory factor)); interferons(such as IFN-γ, IFN-α, and IFN-β); immunoglobin superfamily (such asB7.1 (CD80), and B7.2 (B70, CD86)); TNF family (such as TNF-α(cachectin), TNF-β (lymphotoxin, LT, LT-α), LT-β, CD40 ligand (CD40L),Fas ligand (FasL), CD27 ligand (CD27L), CD30 ligand (CD30L), and4-1BBL)); and those unassigned to a particular family (such as TGF-β,IL1α, IL-1β, IL-1 RA, IL-10 (cytokine synthesis inhibitor F), IL-12 (NKcell stimulatory factor), MIF, IL-16, IL-17 (mCTLA-8), and/or IL-18(IGIF, interferon-γ inducing factor)). Furthermore, the Fc portion ofthe heavy chain of an antibody may be used to target Fcreceptor-expressing cells such as the use of the Fc portion of an IgEantibody to target mast cells and basophils.

Furthermore, in some aspects, the cell-targeting moiety may be a peptidesequence or a cyclic peptide. Examples, cell- and tissue-targetingpeptides that may be used according to the embodiments are provided, forinstance, in U.S. Pat. Nos. 6,232,287; 6,528,481; 7,452,964; 7,671,010;7,781,565; 8,507,445; and 8,450,278, each of which is incorporatedherein by reference.

Thus, in some embodiments, cell targeting moieties are antibodies oravimers. Antibodies and avimers can be generated against virtually anycell surface marker thus, providing a method for targeted to delivery ofGrB to virtually any cell population of interest. Methods for generatingantibodies that may be used as cell targeting moieties are detailedbelow. Methods for generating avimers that bind to a given cell surfacemarker are detailed in U.S. Patent Publications Nos. 2006/0234299 and2006/0223114, each incorporated herein by reference.

V. THERAPIES

A. Pharmaceutical Formulations and Routes of Administration

Where clinical applications are contemplated, it will be necessary toprepare pharmaceutical compositions in a form appropriate for theintended application. In some embodiments, such formulation with thecompounds of the present disclosure is contemplated. Generally, thiswill entail preparing compositions that are essentially free ofpyrogens, as well as other impurities that could be harmful to humans oranimals.

One will generally desire to employ appropriate salts and buffers torender delivery vectors stable and allow for uptake by target cells.Buffers also will be employed when recombinant cells are introduced intoa patient. Aqueous compositions of the present invention comprise aneffective amount of the vector to cells, dissolved or dispersed in apharmaceutically acceptable carrier or aqueous medium. Such compositionsalso are referred to as inocula. The phrase “pharmaceutically orpharmacologically acceptable” refers to molecular entities andcompositions that do not produce adverse, allergic, or other untowardreactions when administered to an animal or a human. As used herein,“pharmaceutically acceptable carrier” includes any and all solvents,dispersion media, coatings, antibacterial and antifungal agents,isotonic and absorption delaying agents and the like. The use of suchmedia and agents for pharmaceutically active substances is well known inthe art. Except insofar as any conventional media or agent isincompatible with the vectors or cells of the present invention, its usein therapeutic compositions is contemplated. Supplementary activeingredients also can be incorporated into the compositions.

The active compositions of the present invention may include classicpharmaceutical preparations. Administration of these compositionsaccording to the present invention will be via any common route so longas the target tissue is available via that route. Such routes includeoral, nasal, buccal, rectal, vaginal or topical route. Alternatively,administration may be by orthotopic, intradermal, subcutaneous,intramuscular, intratumoral, intraperitoneal, or intravenous injection.Such compositions would normally be administered as pharmaceuticallyacceptable compositions, described supra.

The active compounds may also be administered parenterally orintraperitoneally. Solutions of the active compounds as free base orpharmacologically acceptable salts can be prepared in water suitablymixed with a surfactant, such as hydroxypropylcellulose. Dispersions canalso be prepared in glycerol, liquid polyethylene glycols, and mixturesthereof and in oils. Under ordinary conditions of storage and use, thesepreparations contain a preservative to prevent the growth ofmicroorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms, such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), suitable mixtures thereof,and vegetable oils. The proper fluidity can be maintained, for example,by the use of a coating, such as lecithin, by the maintenance of therequired particle size in the case of dispersion and by the use ofsurfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many cases, it will be preferable to include isotonicagents, for example, sugars or sodium chloride. Prolonged absorption ofthe injectable compositions can be brought about by the use in thecompositions of agents delaying absorption, for example, aluminummonostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompounds in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

As used herein, “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents and the like. The use ofsuch media and agents for pharmaceutical active substances is well knownin the art. Except insofar as any conventional media or agent isincompatible with the active ingredient, its use in the therapeuticcompositions is contemplated. Supplementary active ingredients can alsobe incorporated into the compositions.

For oral administration trioxacarcin and derivatives thereof of thepresent disclosure may be incorporated with excipients and used in theform of non-ingestible mouthwashes and dentifrices. A mouthwash may beprepared incorporating the active ingredient in the required amount inan appropriate solvent, such as a sodium borate solution (Dobell'sSolution). Alternatively, the active ingredient may be incorporated intoan antiseptic wash containing sodium borate, glycerin and potassiumbicarbonate. The active ingredient may also be dispersed in dentifrices,including: gels, pastes, powders and slurries. The active ingredient maybe added in a therapeutically effective amount to a paste dentifricethat may include water, binders, abrasives, flavoring agents, foamingagents, and humectants.

The compositions of the present disclosure may be formulated in aneutral or salt form. Pharmaceutically-acceptable salts include the acidaddition salts (formed with the free amino groups of the protein) andwhich are formed with inorganic acids such as, for example, hydrochloricor phosphoric acids, or such organic acids as acetic, oxalic, tartaric,mandelic, and the like. Salts formed with the free carboxyl groups canalso be derived from inorganic bases such as, for example, sodium,potassium, ammonium, calcium, or ferric hydroxides, and such organicbases as isopropylamine, trimethylamine, histidine, procaine and thelike.

Upon formulation, solutions will be administered in a manner compatiblewith the dosage formulation and in such amount as is therapeuticallyeffective. The formulations are easily administered in a variety ofdosage forms such as injectable solutions, drug release capsules and thelike. For parenteral administration in an aqueous solution, for example,the solution should be suitably buffered if necessary and the liquiddiluent first rendered isotonic with sufficient saline or glucose. Theseparticular aqueous solutions are especially suitable for intravenous,intramuscular, subcutaneous and intraperitoneal administration. In thisconnection, sterile aqueous media which can be employed will be known tothose of skill in the art in light of the present disclosure. Forexample, one dosage could be dissolved in 1 ml of isotonic NaCl solutionand either added to 1000 ml of hypodermoclysis fluid or injected at theproposed site of infusion, (see for example, “Remington's PharmaceuticalSciences,” 15th Edition, pages 1035-1038 and 1570-1580). Some variationin dosage will necessarily occur depending on the condition of thesubject being treated. The person responsible for administration will,in any event, determine the appropriate dose for the individual subject.Moreover, for human administration, preparations should meet sterility,pyrogenicity, general safety and purity standards as required by FDAOffice of Biologics standards.

B. Methods of Treatment

In particular, the compositions that may be used in treating microbialinfections and cancer in a subject (e.g., a human subject) are disclosedherein. The compositions described above are preferably administered toa mammal (e.g., rodent, human, non-human primates, canine, bovine,ovine, equine, feline, etc.) in an effective amount, that is, an amountcapable of producing a desirable result in a treated subject (e.g.,causing apoptosis of cancerous cells or killing bacterial cells).Toxicity and therapeutic efficacy of the compositions utilized inmethods of the invention can be determined by standard pharmaceuticalprocedures. As is well known in the medical and veterinary arts, dosagefor any one animal depends on many factors, including the subject'ssize, body surface area, body weight, age, the particular composition tobe administered, time and route of administration, general health, theclinical symptoms of the infection or cancer and other drugs beingadministered concurrently. A composition as described herein istypically administered at a dosage that inhibits the growth orproliferation of a bacterial cell, inhibits the growth of a biofilm, orinduces death of cancerous cells (e.g., induces apoptosis of a cancercell), as assayed by identifying a reduction in hematological parameters(complete blood count—CBC), or cancer cell growth or proliferation. Insome embodiments, amounts of the trioxacarcin derivatives used toinhibit bacterial growth or induce apoptosis of the cancer cells iscalculated to be from about 0.01 mg to about 10,000 mg/day. In someembodiments, the amount is from about 1 mg to about 1,000 mg/day. Insome embodiments, these dosings may be reduced or increased based uponthe biological factors of a particular patient such as increased ordecreased metabolic breakdown of the drug or decreased uptake by thedigestive tract if administered orally. Additionally, the newderivatives of trioxacarcin may be more efficacious and thus a smallerdose is required to achieve a similar effect. Such a dose is typicallyadministered once a day for a few weeks or until sufficient reducing incancer cells has been achieved.

The therapeutic methods of the invention (which include prophylactictreatment) in general include administration of a therapeuticallyeffective amount of the compositions described herein to a subject inneed thereof, including a mammal, particularly a human. Such treatmentwill be suitably administered to subjects, particularly humans,suffering from, having, susceptible to, or at risk for a disease,disorder, or symptom thereof. Determination of those subjects “at risk”can be made by any objective or subjective determination by a diagnostictest or opinion of a subject or health care provider (e.g., genetictest, enzyme or protein marker, marker (as defined herein), familyhistory, and the like).

In one embodiment, the invention provides a method of monitoringtreatment progress. The method includes the step of determining a levelof changes in hematological parameters and/or cancer stem cell (CSC)analysis with cell surface proteins as diagnostic markers (which caninclude, for example, but are not limited to CD34, CD38, CD90, andCD117) or diagnostic measurement (e.g., screen, assay) in a subjectsuffering from or susceptible to a disorder or symptoms thereofassociated with cancer (e.g., leukemia) in which the subject has beenadministered a therapeutic amount of a composition as described herein.The level of marker determined in the method can be compared to knownlevels of marker in either healthy normal controls or in other afflictedpatients to establish the subject's disease status. In preferredembodiments, a second level of marker in the subject is determined at atime point later than the determination of the first level, and the twolevels are compared to monitor the course of disease or the efficacy ofthe therapy. In certain preferred embodiments, a pre-treatment level ofmarker in the subject is determined prior to beginning treatmentaccording to the methods described herein; this pre-treatment level ofmarker can then be compared to the level of marker in the subject afterthe treatment commences, to determine the efficacy of the treatment.

C. Combination Therapies

It is envisioned that the trioxacarcin derivatives described herein maybe used in combination therapies with an additional antimicrobial agentsuch as an antibiotic or a compound which mitigates one or more of theside effects experienced by the patient.

Furthermore, it is very common in the field of cancer therapy to combinetherapeutic modalities. The following is a general discussion oftherapies that may be used in conjunction with the therapies of thepresent disclosure.

To treat cancers using the methods and compositions of the presentdisclosure, one would generally contact a tumor cell or subject with acompound and at least one other therapy. These therapies would beprovided in a combined amount effective to achieve a reduction in one ormore disease parameter. This process may involve contacting thecells/subjects with the both agents/therapies at the same time, e.g.,using a single composition or pharmacological formulation that includesboth agents, or by contacting the cell/subject with two distinctcompositions or formulations, at the same time, wherein one compositionincludes the compound and the other includes the other agent.

Alternatively, trioxacarcin derivatives of the present disclosure mayprecede or follow the other treatment by intervals ranging from minutesto weeks. One would generally ensure that a significant period of timedid not expire between the time of each delivery, such that thetherapies would still be able to exert an advantageously combined effecton the cell/subject. In such instances, it is contemplated that onewould contact the cell with both modalities within about 12-24 hours ofeach other, within about 6-12 hours of each other, or with a delay timeof only about 12 hours. In some situations, it may be desirable toextend the time period for treatment significantly; however, whereseveral days (2, 3, 4, 5, 6 or 7) to several weeks (1, 2, 3, 4, 5, 6, 7or 8) lapse between the respective administrations.

It also is conceivable that more than one administration of either thecompound or the other therapy will be desired. Various combinations maybe employed, where a compound of the present disclosure is “A,” and theother therapy is “B,” as exemplified below:

A/B/A B/A/B B/B/A A/A/B B/A/A A/B/B B/B/B/A B/B/A/B A/A/B/B A/B/A/BA/B/B/A B/B/A/A B/A/B/A B/A/A/B B/B/B/A A/A/A/B B/A/A/A A/B/A/A A/A/B/AA/B/B/B B/A/B/B B/B/A/BAgents or factors suitable for use in a combined therapy with agentsaccording to the present disclosure against an infectious diseaseinclude antibiotics such as penicillins, cephalosporins, carbapenems,macrolides, aminoglycosides, quinolones (including fluoroquinolones),sulfonamides and tetracylcines. Other combinations are contemplated. Thefollowing is a general discussion of antibiotic, antiviral, and cancertherapies that may be used combination with the compounds of the presentdisclosure.

1. Antibiotics

The term “antibiotics” are drugs which may be used to treat a bacterialinfection through either inhibiting the growth of bacteria or killingbacteria. Without being bound by theory, it is believed that antibioticscan be classified into two major classes: bactericidal agents that killbacteria or bacteriostatic agents that slow down or prevent the growthof bacteria.

The first commercially available antibiotic was released in the 1930's.Since then, many different antibiotics have been developed and widelyprescribed. In 2010, on average, 4 in 5 Americans are prescribedantibiotics annually. Given the prevalence of antibiotics, bacteria havestarted to develop resistance to specific antibiotics and antibioticmechanisms. Without being bound by theory, the use of antibiotics incombination with another antibiotic may modulate resistance and enhancethe efficacy of one or both agents.

In some embodiments, antibiotics can fall into a wide range of classes.In some embodiments, the compounds of the present disclosure may be usedin conjunction with another antibiotic. In some embodiments, thecompounds may be used in conjunction with a narrow spectrum antibioticwhich targets a specific bacteria type. In some non-limiting examples ofbactericidal antibiotics include penicillin, cephalosporin, polymyxin,rifamycin, lipiarmycin, quinolones, and sulfonamides. In somenon-limiting examples of bacteriostatic antibiotics include macrolides,lincosamides, or tetracyclines. In some embodiments, the antibiotic isan aminoglycoside such as kanamycin and streptomycin, an ansamycin suchas rifaximin and geldanamycin, a carbacephem such as loracarbef, acarbapenem such as ertapenem, imipenem, a cephalosporin such ascephalexin, cefixime, cefepime, and ceftobiprole, a glycopeptide such asvancomycin or teicoplanin, a lincosamide such as lincomycin andclindamycin, a lipopeptide such as daptomycin, a macrolide such asclarithromycin, spiramycin, azithromycin, and telithromycin, amonobactam such as aztreonam, a nitrofuran such as furazolidone andnitrofurantoin, an oxazolidonones such as linezolid, a penicillin suchas amoxicillin, azlocillin, flucloxacillin, and penicillin G, anantibiotic polypeptide such as bacitracin, polymyxin B, and colistin, aquinolone such as ciprofloxacin, levofloxacin, and gatifloxacin, asulfonamide such as silver sulfadiazine, mefenide, sulfadimethoxine, orsulfasalazine, or a tetracycline such as demeclocycline, doxycycline,minocycline, oxytetracycline, or tetracycline. In some embodiments, thecompounds could be combined with a drug which acts against mycobacteriasuch as cycloserine, capreomycin, ethionamide, rifampicin, rifabutin,rifapentine, and streptomycin. Other antibiotics that are contemplatedfor combination therapies may include arsphenamine, chloramphenicol,fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin,quinupristin, dalfopristin, thiamphenicol, tigecycline, tinidazole, ortrimethoprim.

2. Chemotherapy

The term “chemotherapy” refers to the use of drugs to treat cancer. A“chemotherapeutic agent” is used to connote a compound or compositionthat is administered in the treatment of cancer. These agents or drugsare categorized by their mode of activity within a cell, for example,whether and at what stage they affect the cell cycle. Alternatively, anagent may be characterized based on its ability to directly cross-linkDNA, to intercalate into DNA, or to induce chromosomal and mitoticaberrations by affecting nucleic acid synthesis. Most chemotherapeuticagents fall into the following categories: alkylating agents,antimetabolites, antitumor antibiotics, mitotic inhibitors, andnitrosoureas.

Examples of chemotherapeutic agents include alkylating agents such asthiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan,improsulfan and piposulfan; aziridines such as benzodopa, carboquone,meturedopa, and uredopa; ethylenimines and methylamelamines includingaltretamine, triethylenemelamine, trietylenephosphoramide,triethiylenethiophosphoramide and trimethylolomelamine; acetogenins(especially bullatacin and bullatacinone); a camptothecin (including thesynthetic analogue topotecan); bryostatin; callystatin; CC-1065(including its adozelesin, carzelesin and bizelesin syntheticanalogues); cryptophycins (particularly cryptophycin 1 and cryptophycin8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin;spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine,cholophosphamide, estramustine, ifosfamide, mechlorethamine,mechlorethamine oxide hydrochloride, melphalan, novembichin,phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureassuch as carmustine, chlorozotocin, fotemustine, lomustine, nimustine,and ranimnustine; antibiotics such as the enediyne antibiotics (e.g.,calicheamicin, especially calicheamicin gammalI and calicheamicinomegaI1; dynemicin, including dynemicin A uncialamycin and derivativesthereof; bisphosphonates, such as clodronate; an esperamicin; as well asneocarzinostatin chromophore and related chromoprotein enediyneantiobiotic chromophores, aclacinomysins, actinomycin, authrarnycin,azaserine, bleomycins, cactinomycin, carabicin, carminomycin,carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin (includingmorpholino-doxorubicin, cyanomorpholino-doxorubicin,2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolicacid, nogalarnycin, olivomycins, peplomycin, potfiromycin, puromycin,quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexateand 5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adrenals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as folinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil;amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids suchas maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol;nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK polysaccharidecomplex); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonicacid; triaziquone; 2,2′,2″-trichlorotriethylamine; trichothecenes(especially T-2 toxin, verracurin A, roridin A and anguidine); urethan;vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol;pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide;thiotepa; taxoids, e.g., paclitaxel and docetaxel; chlorambucil;gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinumcoordination complexes such as cisplatin, oxaliplatin and carboplatin;vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone;vincristine; vinorelbine; novantrone; teniposide; edatrexate;daunomycin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11);topoisomerase inhibitor RFS 2000; difluorometlhylornithine (DMFO);retinoids such as retinoic acid; capecitabine; cisplatin (CDDP),carboplatin, procarbazine, mechlorethamine, cyclophosphamide,camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea,dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin,mitomycin, etoposide (VP16), tamoxifen, raloxifene, estrogen receptorbinding agents, taxol, paclitaxel, docetaxel, gemcitabien, navelbine,farnesyl-protein tansferase inhibitors, transplatinum, 5-fluorouracil,vincristin, vinblastin and methotrexate and pharmaceutically acceptablesalts, acids or derivatives of any of the above.

3. Radiotherapy

Radiotherapy, also called radiation therapy, is the treatment of cancerand other diseases with ionizing radiation. Ionizing radiation depositsenergy that injures or destroys cells in the area being treated bydamaging their genetic material, making it impossible for these cells tocontinue to grow. Although radiation damages both cancer cells andnormal cells, the latter are able to repair themselves and functionproperly.

Radiation therapy used according to the present invention may include,but is not limited to, the use of γ-rays, X-rays, and/or the directeddelivery of radioisotopes to tumor cells. Other forms of DNA damagingfactors are also contemplated such as microwaves and UV-irradiation. Itis most likely that all of these factors induce a broad range of damageon DNA, on the precursors of DNA, on the replication and repair of DNA,and on the assembly and maintenance of chromosomes. Dosage ranges forX-rays range from daily doses of 50 to 200 roentgens for prolongedperiods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens.Dosage ranges for radioisotopes vary widely, and depend on the half-lifeof the isotope, the strength and type of radiation emitted, and theuptake by the neoplastic cells.

Radiotherapy may comprise the use of radiolabeled antibodies to deliverdoses of radiation directly to the cancer site (radioimmunotherapy).Antibodies are highly specific proteins that are made by the body inresponse to the presence of antigens (substances recognized as foreignby the immune system). Some tumor cells contain specific antigens thattrigger the production of tumor-specific antibodies. Large quantities ofthese antibodies can be made in the laboratory and attached toradioactive substances (a process known as radiolabeling). Once injectedinto the body, the antibodies actively seek out the cancer cells, whichare destroyed by the cell-killing (cytotoxic) action of the radiation.This approach can minimize the risk of radiation damage to healthycells.

Conformal radiotherapy uses the same radiotherapy machine, a linearaccelerator, as the normal radiotherapy treatment but metal blocks areplaced in the path of the x-ray beam to alter its shape to match that ofthe cancer. This ensures that a higher radiation dose is given to thetumor. Healthy surrounding cells and nearby structures receive a lowerdose of radiation, so the possibility of side effects is reduced. Adevice called a multi-leaf collimator has been developed and may be usedas an alternative to the metal blocks. The multi-leaf collimatorconsists of a number of metal sheets which are fixed to the linearaccelerator. Each layer can be adjusted so that the radiotherapy beamscan be shaped to the treatment area without the need for metal blocks.Precise positioning of the radiotherapy machine is very important forconformal radiotherapy treatment and a special scanning machine may beused to check the position of internal organs at the beginning of eachtreatment.

High-resolution intensity modulated radiotherapy also uses a multi-leafcollimator. During this treatment the layers of the multi-leafcollimator are moved while the treatment is being given. This method islikely to achieve even more precise shaping of the treatment beams andallows the dose of radiotherapy to be constant over the whole treatmentarea.

Although research studies have shown that conformal radiotherapy andintensity modulated radiotherapy may reduce the side effects ofradiotherapy treatment, it is possible that shaping the treatment areaso precisely could stop microscopic cancer cells just outside thetreatment area being destroyed. This means that the risk of the cancercoming back in the future may be higher with these specializedradiotherapy techniques.

Scientists also are looking for ways to increase the effectiveness ofradiation therapy. Two types of investigational drugs are being studiedfor their effect on cells undergoing radiation. Radiosensitizers makethe tumor cells more likely to be damaged, and radioprotectors protectnormal tissues from the effects of radiation. Hyperthermia, the use ofheat, is also being studied for its effectiveness in sensitizing tissueto radiation.

4. Immunotherapy

In the context of cancer treatment, immunotherapeutics, generally, relyon the use of immune effector cells and molecules to target and destroycancer cells. Trastuzumab (Herceptin™) is such an example. The immuneeffector may be, for example, an antibody specific for some marker onthe surface of a tumor cell. The antibody alone may serve as an effectorof therapy or it may recruit other cells to actually affect cellkilling. The antibody also may be conjugated to a drug or toxin(chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussistoxin, etc.) and serve merely as a targeting agent. Alternatively, theeffector may be a lymphocyte carrying a surface molecule that interacts,either directly or indirectly, with a tumor cell target. Variouseffector cells include cytotoxic T cells and NK cells. The combinationof therapeutic modalities, i.e., direct cytotoxic activity andinhibition or reduction of ErbB2 would provide therapeutic benefit inthe treatment of ErbB2 overexpressing cancers.

In one aspect of immunotherapy, the tumor cell must bear some markerthat is amenable to targeting, i.e., is not present on the majority ofother cells. Many tumor markers exist and any of these may be suitablefor targeting in the context of the present invention. Common tumormarkers include carcinoembryonic antigen, prostate specific antigen,urinary tumor associated antigen, fetal antigen, tyrosinase (p97), gp68,TAG-72, HMFG, Sialyl Lewis Antigen, MucA, MucB, PLAP, estrogen receptor,laminin receptor, erb B and p155. An alternative aspect of immunotherapyis to combine anticancer effects with immune stimulatory effects. Immunestimulating molecules also exist including: cytokines such as IL-2,IL-4, IL-12, GM-CSF, γ-IFN, chemokines such as MIP-1, MCP-1, IL-8 andgrowth factors such as FLT3 ligand. Combining immune stimulatingmolecules, either as proteins or using gene delivery in combination witha tumor suppressor has been shown to enhance antitumor effects (Ju etal., 2000). Moreover, antibodies against any of these compounds may beused to target the anti-cancer agents discussed herein.

Examples of immunotherapies currently under investigation or in use areimmune adjuvants e.g., Mycobacterium bovis, Plasmodium falciparum,dinitrochlorobenzene and aromatic compounds (U.S. Pat. Nos. 5,801,005and 5,739,169; Hui and Hashimoto, 1998; Christodoulides et al., 1998),cytokine therapy, e.g., interferons α, β, and γ; IL-1, GM-CSF and TNF(Bukowski et al., 1998; Davidson et al., 1998; Hellstrand et al., 1998)gene therapy, e.g., TNF, IL-1, IL-2, p53 (Qin et al., 1998; Austin-Wardand Villaseca, 1998; U.S. Pat. Nos. 5,830,880 and 5,846,945) andmonoclonal antibodies, e.g., anti-ganglioside GM2, anti-HER-2, anti-p185(Pietras et al., 1998; Hanibuchi et al., 1998; U.S. Pat. No. 5,824,311).It is contemplated that one or more anti-cancer therapies may beemployed with the gene silencing therapies described herein.

In active immunotherapy, an antigenic peptide, polypeptide or protein,or an autologous or allogenic tumor cell composition or “vaccine” isadministered, generally with a distinct bacterial adjuvant (Ravindranathand Morton, 1991; Morton et al., 1992; Mitchell et al., 1990; Mitchellet al., 1993).

In adoptive immunotherapy, the patient's circulating lymphocytes, ortumor infiltrated lymphocytes, are isolated in vitro, activated bylymphokines such as IL-2 or transduced with genes for tumor necrosis,and readministered (Rosenberg et al., 1988; 1989).

5. Surgery

Approximately 60% of persons with cancer will undergo surgery of sometype, which includes preventative, diagnostic or staging, curative, andpalliative surgery. Curative surgery is a cancer treatment that may beused in conjunction with other therapies, such as the treatment of thepresent invention, chemotherapy, radiotherapy, hormonal therapy, genetherapy, immunotherapy and/or alternative therapies.

Curative surgery includes resection in which all or part of canceroustissue is physically removed, excised, and/or destroyed. Tumor resectionrefers to physical removal of at least part of a tumor. In addition totumor resection, treatment by surgery includes laser surgery,cryosurgery, electrosurgery, and microscopically controlled surgery(Mohs' surgery). It is further contemplated that the present inventionmay be used in conjunction with removal of superficial cancers,precancers, or incidental amounts of normal tissue.

Upon excision of part or all of cancerous cells, tissue, or tumor, acavity may be formed in the body. Treatment may be accomplished byperfusion, direct injection or local application of the area with anadditional anti-cancer therapy. Such treatment may be repeated, forexample, every 1, 2, 3, 4, 5, 6, or 7 days, or every 1, 2, 3, 4, and 5weeks or every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. Thesetreatments may be of varying dosages as well.

In some particular embodiments, after removal of the tumor, an adjuvanttreatment with a compound of the present disclosure is believe to beparticularly efficacious in reducing the reoccurrence of the tumor.Additionally, the compounds of the present disclosure can also be usedin a neoadjuvant setting.

6. Other Agents

It is contemplated that other agents may be used with the presentinvention. These additional agents include immunomodulatory agents,agents that affect the upregulation of cell surface receptors and GAPjunctions, cytostatic and differentiation agents, inhibitors of celladhesion, agents that increase the sensitivity of the hyperproliferativecells to apoptotic inducers, or other biological agents.Immunomodulatory agents include tumor necrosis factor; interferon alpha,beta, and gamma; IL-2 and other cytokines; F42K and other cytokineanalogs; or MIP-1, MIP-1β, MCP-1, RANTES, and other chemokines. It isfurther contemplated that the upregulation of cell surface receptors ortheir ligands such as Fas/Fas ligand, DR4 or DR5/TRAIL (Apo-2 ligand)would potentiate the apoptotic inducing abilities of the presentinvention by establishment of an autocrine or paracrine effect onhyperproliferative cells. Increases intercellular signaling by elevatingthe number of GAP junctions would increase the anti-hyperproliferativeeffects on the neighboring hyperproliferative cell population. In otherembodiments, cytostatic or differentiation agents may be used incombination with the present invention to improve theanti-hyperproliferative efficacy of the treatments. Inhibitors of celladhesion are contemplated to improve the efficacy of the presentinvention. Examples of cell adhesion inhibitors are focal adhesionkinase (FAKs) inhibitors and Lovastatin. It is further contemplated thatother agents that increase the sensitivity of a hyperproliferative cellto apoptosis, such as the antibody c225, could be used in combinationwith the present invention to improve the treatment efficacy.

There have been many advances in the therapy of cancer following theintroduction of cytotoxic chemotherapeutic drugs. However, one of theconsequences of chemotherapy is the development/acquisition ofdrug-resistant phenotypes and the development of multiple drugresistance. The development of drug resistance remains a major obstaclein the treatment of such tumors and therefore, there is an obvious needfor alternative approaches such as gene therapy.

Another form of therapy for use in conjunction with chemotherapy,radiation therapy or biological therapy includes hyperthermia, which isa procedure in which a patient's tissue is exposed to high temperatures(up to 106° F.). External or internal heating devices may be involved inthe application of local, regional, or whole-body hyperthermia. Localhyperthermia involves the application of heat to a small area, such as atumor. Heat may be generated externally with high-frequency wavestargeting a tumor from a device outside the body. Internal heat mayinvolve a sterile probe, including thin, heated wires or hollow tubesfilled with warm water, implanted microwave antennae, or radiofrequencyelectrodes.

A patient's organ or a limb is heated for regional therapy, which isaccomplished using devices that produce high energy, such as magnets.Alternatively, some of the patient's blood may be removed and heatedbefore being perfused into an area that will be internally heated.Whole-body heating may also be implemented in cases where cancer hasspread throughout the body. Warm-water blankets, hot wax, inductivecoils, and thermal chambers may be used for this purpose.

The skilled artisan is directed to “Remington's Pharmaceutical Sciences”15th Edition, chapter 33, in particular pages 624-652. Some variation indosage will necessarily occur depending on the condition of the subjectbeing treated. The person responsible for administration will, in anyevent, determine the appropriate dose for the individual subject.Moreover, for human administration, preparations should meet sterility,pyrogenicity, general safety and purity standards as required by FDAOffice of Biologics standards.

It also should be pointed out that any of the foregoing therapies mayprove useful by themselves in treating cancer.

VI. Synthetic Methods

In some aspects, the compounds of this invention can be synthesizedusing the methods of organic chemistry as described in this application.These methods can be further modified and optimized using the principlesand techniques of organic chemistry as applied by a person skilled inthe art. Such principles and techniques are taught, for example, inMarch's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure(2007), which is incorporated by reference herein.

A. Process Scale-Up

The synthetic methods described herein can be further modified andoptimized for preparative, pilot- or large-scale production, eitherbatch of continuous, using the principles and techniques of processchemistry as applied by a person skilled in the art. Such principles andtechniques are taught, for example, in Practical Process Research &Development (2000), which is incorporated by reference herein. Thesynthetic method described herein may be used to produce preparativescale amounts of trioxacarcin and derivatives thereof.

B. Chemical Definitions

When used in the context of a chemical group: “hydrogen” means —H;“hydroxy” means —OH; “oxo” means ═O; “carbonyl” means —C(═O)—; “carboxy”means —C(═O)OH (also written as —COOH or —CO₂H); “halo” meansindependently —F, —Cl, —Br or —I; “amino” means —NH₂; “hydroxyamino”means —NHOH; “nitro” means —NO₂; imino means ═NH; “cyano” means —CN;“isocyanate” means —N═C═O; “azido” means —N₃; in a monovalent context“phosphate” means —OP(O)(OH)₂ or a deprotonated form thereof; in adivalent context “phosphate” means —OP(O)(OH)O— or a deprotonated formthereof; “mercapto” means —SH; and “thio” means ═S; “sulfato” means—SO₃H, “sulfamido” means —S(O)₂NH₂, “sulfonyl” means —S(O)₂—; and“sulfinyl” means —S(O)—.

In the context of chemical formulas, the symbol “—” means a single bond,“═” means a double bond, and “≡” means triple bond. The symbol “----”represents an optional bond, which if present is either single ordouble. The symbol “

” represents a single bond or a double bond. Thus, for example, theformula

includes

And it is understood that no one such ring atom forms part of more thanone double bond. Furthermore, it is noted that the covalent bond symbol“—”, when connecting one or two stereogenic atoms, does not indicate anypreferred stereochemistry. Instead, it covers all stereoisomers as wellas mixtures thereof. The symbol “

”, when drawn perpendicularly across a bond (e.g.,

for methyl) indicates a point of attachment of the group. It is notedthat the point of attachment is typically only identified in this mannerfor larger groups in order to assist the reader in unambiguouslyidentifying a point of attachment. The symbol “

” means a single bond where the group attached to the thick end of thewedge is “out of the page.” The symbol “

” means a single bond where the group attached to the thick end of thewedge is “into the page”. The symbol “

” means a single bond where the geometry around a double bond (e.g.,either E or Z) is undefined. Both options, as well as combinationsthereof are therefore intended. Any undefined valency on an atom of astructure shown in this application implicitly represents a hydrogenatom bonded to that atom. A bold dot on a carbon atom indicates that thehydrogen attached to that carbon is oriented out of the plane of thepaper.

When a group “R” is depicted as a “floating group” on a ring system, forexample, in the formula:

then R may replace any hydrogen atom attached to any of the ring atoms,including a depicted, implied, or expressly defined hydrogen, so long asa stable structure is formed. When a group “R” is depicted as a“floating group” on a fused ring system, as for example in the formula:

then R may replace any hydrogen attached to any of the ring atoms ofeither of the fused rings unless specified otherwise. Replaceablehydrogens include depicted hydrogens (e.g., the hydrogen attached to thenitrogen in the formula above), implied hydrogens (e.g., a hydrogen ofthe formula above that is not shown but understood to be present),expressly defined hydrogens, and optional hydrogens whose presencedepends on the identity of a ring atom (e.g., a hydrogen attached togroup X, when X equals —CH—), so long as a stable structure is formed.In the example depicted, R may reside on either the 5-membered or the6-membered ring of the fused ring system. In the formula above, thesubscript letter “y” immediately following the group “R” enclosed inparentheses, represents a numeric variable. Unless specified otherwise,this variable can be 0, 1, 2, or any integer greater than 2, onlylimited by the maximum number of replaceable hydrogen atoms of the ringor ring system.

For the groups and classes below, the following parenthetical subscriptsfurther define the group/class as follows: “(Cn)” defines the exactnumber (n) of carbon atoms in the group/class. “(Cn)” defines themaximum number (n) of carbon atoms that can be in the group/class, withthe minimum number as small as possible for the group in question, e.g.,it is understood that the minimum number of carbon atoms in the group“alkenyl_((C≤8))” or the class “alkene_((C≤8))” is two. For example,“alkoxy_((C≤10))” designates those alkoxy groups having from 1 to 10carbon atoms. (Cn-n′) defines both the minimum (n) and maximum number(n′) of carbon atoms in the group. Similarly, “alkyl_((C2-10))”designates those alkyl groups having from 2 to 10 carbon atoms.

The term “saturated” as used herein means the compound or group somodified has no carbon-carbon double and no carbon-carbon triple bonds,except as noted below. In the case of substituted versions of saturatedgroups, one or more carbon oxygen double bond or a carbon nitrogendouble bond may be present. And when such a bond is present, thencarbon-carbon double bonds that may occur as part of keto-enoltautomerism or imine/enamine tautomerism are not precluded.

The term “aliphatic” when used without the “substituted” modifiersignifies that the compound/group so modified is an acyclic or cyclic,but non-aromatic hydrocarbon compound or group. In aliphaticcompounds/groups, the carbon atoms can be joined together in straightchains, branched chains, or non-aromatic rings (alicyclic). Aliphaticcompounds/groups can be saturated, that is joined by single bonds(alkanes/alkyl), or unsaturated, with one or more double bonds(alkenes/alkenyl) or with one or more triple bonds (alkynes/alkynyl).

The term “alkyl” when used without the “substituted” modifier refers toa monovalent saturated aliphatic group with a carbon atom as the pointof attachment, a linear or branched acyclic structure, and no atomsother than carbon and hydrogen. The groups —CH₃ (Me), —CH₂CH₃ (Et),—CH₂CH₂CH₃ (n-Pr or propyl), —CH(CH₃)₂ (i-Pr, ^(i)Pr or isopropyl),—CH₂CH₂CH₂CH₃ (n-Bu), —CH(CH₃)CH₂CH₃ (sec-butyl), —CH₂CH(CH₃)₂(isobutyl), —C(CH₃)₃ (tert-butyl, t-butyl, t-Bu or ^(t)Bu), and—CH₂C(CH₃)₃ (neo-pentyl) are non-limiting examples of alkyl groups. Theterm “alkanediyl” when used without the “substituted” modifier refers toa divalent saturated aliphatic group, with one or two saturated carbonatom(s) as the point(s) of attachment, a linear or branched acyclicstructure, no carbon-carbon double or triple bonds, and no atoms otherthan carbon and hydrogen. The groups, —CH₂— (methylene), —CH₂CH₂—,—CH₂C(CH₃)₂CH₂—, and —CH₂CH₂CH₂—, are non-limiting examples ofalkanediyl groups. The term “alkylidene” when used without the“substituted” modifier refers to the divalent group ═CRR′ in which R andR′ are independently hydrogen or alkyl. Non-limiting examples ofalkylidene groups include: ═CH₂, ═CH(CH₂CH₃), and ═C(CH₃)₂. An “alkane”refers to the compound H—R, wherein R is alkyl as this term is definedabove. When any of these terms is used with the “substituted” modifierone or more hydrogen atom has been independently replaced by —OH, —F,—Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃,—C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. The following groups are non-limiting examples of substitutedalkyl groups: —CH₂OH, —CH₂Cl, —CF₃, —CH₂CN, —CH₂C(O)OH, CH₂C(O)OCH₃,CH₂C(O)NH₂, CH₂C(O)CH₃, —CH₂OCH₃, CH₂OC(O)CH₃, CH₂NH₂, —CH₂N(CH₃)₂, and—CH₂CH₂Cl. The term “haloalkyl” is a subset of substituted alkyl, inwhich one or more hydrogen atoms has been substituted with a halo groupand no other atoms aside from carbon, hydrogen and halogen are present.The group, —CH₂Cl is a non-limiting example of a haloalkyl. The term“fluoroalkyl” is a subset of substituted alkyl, in which one or morehydrogen has been substituted with a fluoro group and no other atomsaside from carbon, hydrogen and fluorine are present. The groups, —CH₂F,—CF₃, and —CH₂CF₃ are non-limiting examples of fluoroalkyl groups.

The term “cycloalkyl” when used without the “substituted” modifierrefers to a monovalent saturated aliphatic group with a carbon atom asthe point of attachment, said carbon atom forms part of one or morenon-aromatic ring structures, a cyclo or cyclic structure, nocarbon-carbon double or triple bonds, and no atoms other than carbon andhydrogen. Non-limiting examples of cycloalkyl groups include: —CH(CH₂)₂(cyclopropyl), cyclobutyl, cyclopentyl, or cyclohexyl. The term“cycloalkanediyl” when used without the “substituted” modifier refers toa divalent saturated aliphatic group with one or two carbon atom as thepoint(s) of attachment, said carbon atom(s) forms part of one or morenon-aromatic ring structures, a cyclo or cyclic structure, nocarbon-carbon double or triple bonds, and no atoms other than carbon andhydrogen.

are non-limiting examples of cycloalkanediyl groups. A “cycloalkane”refers to the compound H—R, wherein R is cycloalkyl as this term isdefined above. When any of these terms is used with the “substituted”modifier one or more hydrogen atom has been independently replaced by—OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃,—OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. The following groups are non-limiting examples of substitutedcycloalkyl groups: —C(OH)(CH₂)₂,

The term “alkenyl” when used without the “substituted” modifier refersto a monovalent unsaturated aliphatic group with a carbon atom as thepoint of attachment, a linear or branched, acyclic structure, at leastone nonaromatic carbon-carbon double bond, no carbon-carbon triplebonds, and no atoms other than carbon and hydrogen. Non-limitingexamples of alkenyl groups include: —CH═CH₂ (vinyl), —CH═CHCH₃,—CH═CHCH₂CH₃, —CH₂CH═CH₂ (allyl), —CH₂CH═CHCH₃, and —CH═CHCH═CH₂. Theterm “alkenediyl” when used without the “substituted” modifier refers toa divalent unsaturated aliphatic group, with two carbon atoms as pointsof attachment, a linear or branched, cyclo, cyclic or acyclic structure,at least one nonaromatic carbon-carbon double bond, no carbon-carbontriple bonds, and no atoms other than carbon and hydrogen. The groups,—CH═CH—, —CH═C(CH₃)CH₂—, and —CH═CHCH₂—, are non-limiting examples ofalkenediyl groups. It is noted that while the alkenediyl group isaliphatic, once connected at both ends, this group is not precluded fromforming part of an aromatic structure. The terms “alkene” and refer to acompound having the formula H—R, wherein R is alkenyl as this term isdefined above. A “terminal alkene” refers to an alkene having just onecarbon-carbon double bond, wherein that bond forms a vinyl group at oneend of the molecule. When any of these terms are used with the“substituted” modifier one or more hydrogen atom has been independentlyreplaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN,—SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂,—OC(O)CH₃, or —S(O)₂NH₂. The groups, —CH═CHF, —CH═CHCl and —CH═CHBr, arenon-limiting examples of substituted alkenyl groups.

The term “cycloalkenyl” when used without the “substituted” modifierrefers to a monovalent unsaturated aliphatic group with a carbon atom asthe point of attachment, said carbon atom forms part of one or morenon-aromatic ring structures, a cyclo or cyclic structure, at least onenon-aromatic carbon-carbon double bond, no carbon-carbon triple bonds,and no atoms other than carbon and hydrogen. In some non-limitingexamples of cycloalkenyl groups include

The term “cycloalkenediyl” when used without the “substituted” modifierrefers to a divalent unsaturated aliphatic group with one or two carbonatom(s) as the point(s) of attachment, said carbon atom(s) forms part ofone or more non-aromatic ring structures, a cyclo or cyclic structure,at least one non-aromatic carbon-carbon double bond, no carbon-carbontriple bonds, and no atoms other than carbon and hydrogen.

are non-limiting examples of cycloalkenediyl. It is noted that while thecycloalkenediyl group is aliphatic, once connected at both ends, thisgroup is not precluded from forming part of an aromatic structure. Theterms “cycloalkene” and refer to a compound having the formula H—R,wherein R is cycloalkenyl as this term is defined above. The term“olefin” is synonymous with the terms “alkene” or a “cycloalkane” asthose terms are defined above. When any of these terms are used with the“substituted” modifier one or more hydrogen atom has been independentlyreplaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN,—SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂,—OC(O)CH₃, or —S(O)₂NH₂. In some non-limiting examples of substitutedcycloalkenyl include

The term “alkynyl” when used without the “substituted” modifier refersto a monovalent unsaturated aliphatic group with a carbon atom as thepoint of attachment, a linear or branched, acyclic structure, at leastone carbon-carbon triple bond, and no atoms other than carbon andhydrogen. As used herein, the term alkynyl does not preclude thepresence of one or more non-aromatic carbon-carbon double bonds. Thegroups, —C≡CH, —C≡CCH₃, and —CH₂C≡CCH₃, are non-limiting examples ofalkynyl groups. An “alkyne” refers to the compound H—R, wherein R isalkynyl. When any of these terms are used with the “substituted”modifier one or more hydrogen atom has been independently replaced by—OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃,—OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂.

The term “aryl” when used without the “substituted” modifier refers to amonovalent unsaturated aromatic group with an aromatic carbon atom asthe point of attachment, said carbon atom forming part of a one or moresix-membered aromatic ring structure, wherein the ring atoms are allcarbon, and wherein the group consists of no atoms other than carbon andhydrogen. If more than one ring is present, the rings may be fused orunfused. As used herein, the term does not preclude the presence of oneor more alkyl or aralkyl groups (carbon number limitation permitting)attached to the first aromatic ring or any additional aromatic ringpresent. Non-limiting examples of aryl groups include phenyl (Ph),methylphenyl, (dimethyl)phenyl, —C₆H₄CH₂CH₃ (ethylphenyl), naphthyl, anda monovalent group derived from biphenyl. The term “arenediyl” when usedwithout the “substituted” modifier refers to a divalent aromatic groupwith two aromatic carbon atoms as points of attachment, said carbonatoms forming part of one or more six-membered aromatic ringstructure(s) wherein the ring atoms are all carbon, and wherein themonovalent group consists of no atoms other than carbon and hydrogen. Asused herein, the term does not preclude the presence of one or morealkyl, aryl or aralkyl groups (carbon number limitation permitting)attached to the first aromatic ring or any additional aromatic ringpresent. If more than one ring is present, the rings may be fused orunfused. Unfused rings may be connected via one or more of thefollowing: a covalent bond, alkanediyl, or alkenediyl groups (carbonnumber limitation permitting). Non-limiting examples of arenediyl groupsinclude:

An “arene” refers to the compound H—R, wherein R is aryl as that term isdefined above. Benzene and toluene are non-limiting examples of arenes.When any of these terms are used with the “substituted” modifier one ormore hydrogen atom has been independently replaced by —OH, —F, —Cl, —Br,—I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃,—C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂.

The term “aralkyl” when used without the “substituted” modifier refersto the monovalent group -alkanediyl-aryl, in which the terms alkanediyland aryl are each used in a manner consistent with the definitionsprovided above. Non-limiting examples of aralkyls are: phenylmethyl(benzyl, Bn) and 2-phenyl-ethyl. When the term aralkyl is used with the“substituted” modifier one or more hydrogen atom from the alkanediyland/or the aryl group has been independently replaced by —OH, —F, —Cl,—Br, —I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃,—C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. Non-limiting examples of substituted aralkyls are:(3-chlorophenyl)-methyl, and 2-chloro-2-phenyl-eth-1-yl.

The term “heteroaryl” when used without the “substituted” modifierrefers to a monovalent aromatic group with an aromatic carbon atom ornitrogen atom as the point of attachment, said carbon atom or nitrogenatom forming part of one or more aromatic ring structures wherein atleast one of the ring atoms is nitrogen, oxygen or sulfur, and whereinthe heteroaryl group consists of no atoms other than carbon, hydrogen,aromatic nitrogen, aromatic oxygen and aromatic sulfur. If more than onering is present, the rings may be fused or unfused. As used herein, theterm does not preclude the presence of one or more alkyl, aryl, and/oraralkyl groups (carbon number limitation permitting) attached to thearomatic ring or aromatic ring system. Non-limiting examples ofheteroaryl groups include furanyl, imidazolyl, indolyl, indazolyl,isoxazolyl, methylpyridinyl, oxazolyl, phenylpyridinyl, pyridinyl,pyrrolyl, pyrimidinyl, pyrazinyl, quinolyl, quinazolyl, quinoxalinyl,triazinyl, tetrazolyl, thiazolyl, thienyl, and triazolyl. The term“N-heteroaryl” refers to a heteroaryl group with a nitrogen atom as thepoint of attachment. The term “heteroarenediyl” when used without the“substituted” modifier refers to an divalent aromatic group, with twoaromatic carbon atoms, two aromatic nitrogen atoms, or one aromaticcarbon atom and one aromatic nitrogen atom as the two points ofattachment, said atoms forming part of one or more aromatic ringstructure(s) wherein at least one of the ring atoms is nitrogen, oxygenor sulfur, and wherein the divalent group consists of no atoms otherthan carbon, hydrogen, aromatic nitrogen, aromatic oxygen and aromaticsulfur. If more than one ring is present, the rings may be fused orunfused. Unfused rings may be connected via one or more of thefollowing: a covalent bond, alkanediyl, or alkenediyl groups (carbonnumber limitation permitting). As used herein, the term does notpreclude the presence of one or more alkyl, aryl, and/or aralkyl groups(carbon number limitation permitting) attached to the aromatic ring oraromatic ring system. Non-limiting examples of heteroarenediyl groupsinclude:

A “heteroarene” refers to the compound H—R, wherein R is heteroaryl.Pyridine and quinoline are non-limiting examples of heteroarenes. Whenthese terms are used with the “substituted” modifier one or morehydrogen atom has been independently replaced by —OH, —F, —Cl, —Br, —I,—NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃,—NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂.

The term “heteroaralkyl” when used without the “substituted” modifierrefers to the monovalent group -alkanediyl-heteroaryl, in which theterms alkanediyl and heteroaryl are each used in a manner consistentwith the definitions provided above. Non-limiting examples ofheteroaralkyls are: 2-pyridylmethyl and 2-indazolyl-ethyl. When the termheteroaralkyl is used with the “substituted” modifier one or morehydrogen atom from the alkanediyl and/or the heteroaryl group has beenindependently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —N₃, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂. Non-limiting examples ofsubstituted heteroaralkyls are: (3-chloroquinolyl)-methyl, and2-chloro-2-thienyl-eth-1-yl.

The term “heterocycloalkyl” when used without the “substituted” modifierrefers to a monovalent non-aromatic group with a carbon atom or nitrogenatom as the point of attachment, said carbon atom or nitrogen atomforming part of one or more non-aromatic ring structures wherein atleast one of the ring atoms is nitrogen, oxygen or sulfur, and whereinthe heterocycloalkyl group consists of no atoms other than carbon,hydrogen, nitrogen, oxygen and sulfur. If more than one ring is present,the rings may be fused or unfused. As used herein, the term does notpreclude the presence of one or more alkyl groups (carbon numberlimitation permitting) attached to the ring or ring system. Also, theterm does not preclude the presence of one or more double bonds in thering or ring system, provided that the resulting group remainsnon-aromatic. Non-limiting examples of heterocycloalkyl groups includeaziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydrothiofuranyl,tetrahydropyranyl, pyranyl, oxiranyl, and oxetanyl. The term“N-heterocycloalkyl” refers to a heterocycloalkyl group with a nitrogenatom as the point of attachment. The term “heterocycloalkanediyl” whenused without the “substituted” modifier refers to an divalent cyclicgroup, with two carbon atoms, two nitrogen atoms, or one carbon atom andone nitrogen atom as the two points of attachment, said atoms formingpart of one or more ring structure(s) wherein at least one of the ringatoms is nitrogen, oxygen or sulfur, and wherein the divalent groupconsists of no atoms other than carbon, hydrogen, nitrogen, oxygen andsulfur. If more than one ring is present, the rings may be fused orunfused. Unfused rings may be connected via one or more of thefollowing: a covalent bond, alkanediyl, or alkenediyl groups (carbonnumber limitation permitting). As used herein, the term does notpreclude the presence of one or more alkyl groups (carbon numberlimitation permitting) attached to the ring or ring system. Also, theterm does not preclude the presence of one or more double bonds in thering or ring system, provided that the resulting group remainsnon-aromatic. Non-limiting examples of heterocycloalkanediyl groupsinclude:

When these terms are used with the “substituted” modifier one or morehydrogen atom has been independently replaced by —OH, —F, —Cl, —Br, —I,—NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃,—NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, —S(O)₂NH₂, or—C(O)OC(CH₃)₃ (tert-butyloxycarbonyl, BOC).

The term “acyl” when used without the “substituted” modifier refers tothe group —C(O)R, in which R is a hydrogen, alkyl, cycloalkyl, aryl,aralkyl or heteroaryl, as those terms are defined above. The groups,—CHO, —C(O)CH₃ (acetyl, Ac), —C(O)CH₂CH₃, —C(O)CH₂CH₂CH₃, —C(O)CH(CH₃)₂,C(O)CH(CH₂)₂, C(O)C₆H₅, —C(O)C₆H₄CH₃, —C(O)CH₂C₆H₅, —C(O)(imidazolyl)are non-limiting examples of acyl groups. A “thioacyl” is defined in ananalogous manner, except that the oxygen atom of the group —C(O)R hasbeen replaced with a sulfur atom, —C(S)R. The term “aldehyde”corresponds to an alkane, as defined above, wherein at least one of thehydrogen atoms has been replaced with a —CHO group. When any of theseterms are used with the “substituted” modifier one or more hydrogen atom(including a hydrogen atom directly attached the carbonyl orthiocarbonyl group, if any) has been independently replaced by —OH, —F,—Cl, —Br, —I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃,—OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂. The groups, —C(O)CH₂CF₃, —CO₂H (carboxyl), —CO₂CH₃(methylcarboxyl), —CO₂CH₂CH₃, —C(O)NH₂ (carbamoyl), and —CON(CH₃)₂, arenon-limiting examples of substituted acyl groups.

The term “alkylamino” when used without the “substituted” modifierrefers to the group —NHR, in which R is an alkyl, as that term isdefined above. Non-limiting examples of alkylamino groups include:—NHCH₃ and —NHCH₂CH₃. The term “dialkylamino” when used without the“substituted” modifier refers to the group —NRR′, in which R and R′ caneach independently be the same or different alkyl groups, or R and R′can be taken together to represent an alkanediyl. Non-limiting examplesof dialkylamino groups include: —N(CH₃)₂, —N(CH₃)(CH₂CH₃), andN-pyrrolidinyl. The terms “alkoxyamino”, “cycloalkylamino”,“alkenylamino”, “cycloalkenylamino”, “alkynylamino”, “arylamino”,“aralkylamino”, “heteroarylamino”, “heterocycloalkylamino” and“alkylsulfonylamino” when used without the “substituted” modifier,refers to groups, defined as —NHR, in which R is alkoxy, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, heteroaryl,heterocycloalkyl, and alkylsulfonyl, respectively. A non-limitingexample of an arylamino group is —NHC₆H₅. The term “amido” (acylamino),when used without the “substituted” modifier, refers to the group —NHR,in which R is acyl, as that term is defined above. A non-limitingexample of an amido group is —NHC(O)CH₃. The term “alkylimino” when usedwithout the “substituted” modifier refers to the divalent group ═NR, inwhich R is an alkyl, as that term is defined above. The term“alkylaminodiyl” refers to the divalent group —NH-alkanediyl-,—NH-alkanediyl-NH—, or -alkanediyl-NH-alkanediyl-. When any of theseterms is used with the “substituted” modifier one or more hydrogen atomhas been independently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂,—N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃,—NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or —S(O)₂NH₂. The groups—NHC(O)OCH₃ and —NHC(O)NHCH₃ are non-limiting examples of substitutedamido groups.

The term “alkoxy” when used without the “substituted” modifier refers tothe group —OR, in which R is an alkyl, as that term is defined above.Non-limiting examples include: —OCH₃ (methoxy), —OCH₂CH₃ (ethoxy),—OCH₂CH₂CH₃, —OCH(CH₃)₂ (isopropoxy), and —OC(CH₃)₃ (tert-butoxy). Theterms “cycloalkoxy”, “alkenyloxy”, “alkynyloxy”, “aryloxy”, “aralkoxy”,“heteroaryloxy”, “heterocycloalkoxy”, and “acyloxy”, when used withoutthe “substituted” modifier, refers to groups, defined as —OR, in which Ris cycloalkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,heterocycloalkyl, and acyl, respectively. The term “alkoxydiyl” refersto the divalent group —O-alkanediyl-, —O-alkanediyl-O—, or-alkanediyl-O-alkanediyl-. The term “alkylthio” and “acylthio” when usedwithout the “substituted” modifier refers to the group —SR, in which Ris an alkyl and acyl, respectively. The term “alcohol” corresponds to analkane, as defined above, wherein at least one of the hydrogen atoms hasbeen replaced with a hydroxy group. The term “ether” corresponds to analkane or cycloalkane, as defined above, wherein at least one of thehydrogen atoms has been replaced with an alkoxy or cycloalkoxy group.When any of these terms is used with the “substituted” modifier one ormore hydrogen atom has been independently replaced by —OH, —F, —Cl, —Br,—I, —NH₂, —NO₂, —N₃, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃,—C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —OC(O)CH₃, or—S(O)₂NH₂.

A “base” in the context of this application is a compound which has alone pair of electron that can accept a proton. Non-limiting examples ofa base can include triethylamine, a metal hydroxide, a metal alkoxide, ametal hydride, or a metal alkane. An alkyllithium or organolithium is acompound of the formula alkyl_((C≤12))-Li. A nitrogenous base is analkylamine, dialkylamino, trialkylamine, nitrogen containingheterocycloalkane or heteroarene wherein the base can accept a proton toform a positively charged species. For example, but not limited to, anitrogenous base could be 4,4-dimethylpyridine, pyridine,1,8-diazabicyclo[5.4.0]undec-7-ene, diisopropylethylamine, ortriethylamine. A metal alkoxide is an alkoxy group wherein the oxygenatom, which was the point of connectivity, has an extra electron andthus a negative charge which is charged balanced by the metal ion. Forexample, a metal alkoxide could be a sodium tert-butoxide or potassiummethoxide.

An “oxidizing agent” in the context of this application is a compoundwhich causes the oxidation of a compound by accepting an electron. Somenon-limiting examples of oxidizing agent are oxygen gas, peroxides,chlorite, hypochlorite, or a chromium compound such as pyridiniumchlorochromate or hydrochromic acid.

A “metal” in the context of this application is a transition metal or ametal of groups I or II. It may also be an element of Group 13 such as,but not limited to, boron and aluminum.

A “Lewis acid” is a atom or functional group which can accept a pair ofelectrons. In some embodiments, the Lewis acid is a metal atom. Withoutbeing bound by any theory, the Lewis acid increases the reactivity ofone or more group to which it attached by increasing the polarization ofa bond.

A “linker” in the context of this application is divalent chemical groupwhich may be used to join one or more molecules to the compound of theinstant disclosure. In some embodiments, the linker contains a reactivefunctional group, such as a carboxyl, an amide, a amine, a hydroxy, amercapto, an aldehyde, or a ketone on each end that be used to join oneor more molecules to the compounds of the instant disclosure. In somenon-limiting examples, —CH₂CH₂CH₂CH₂—, —C(O)CH₂CH₂CH₂—, —OCH₂CH₂NH—,—NHCH₂CH₂NH—, and —(OCH₂CH₂)_(n)— wherein n is between 1-1000, arelinkers.

An “amine protecting group” is well understood in the art. An amineprotecting group is a group which prevents the reactivity of the aminegroup during a reaction which modifies some other portion of themolecule and can be easily removed to generate the desired amine. Amineprotecting groups can be found at least in Greene and Wuts, 1999, whichis incorporated herein by reference. Some non-limiting examples of aminoprotecting groups include formyl, acetyl, propionyl, pivaloyl,t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl,trichloroacetyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl,4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and the like; sulfonylgroups such as benzenesulfonyl, p-toluenesulfonyl and the like; alkoxy-or aryloxycarbonyl groups (which form urethanes with the protectedamine) such as benzyloxycarbonyl (Cbz), p-chlorobenzyloxycarbonyl,p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl,2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl,3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl,2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl,2-nitro-4,5-dimethoxybenzyloxycarbonyl,3,4,5-trimethoxybenzyloxycarbonyl,1-(p-biphenylyl)-1-methylethoxycarbonyl,α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl,t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl,isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl(Alloc), 2,2,2-trichloroethoxycarbonyl, 2-trimethylsilylethyloxycarbonyl(Teoc), phenoxycarbonyl, 4-nitrophenoxycarbonyl,fluorenyl-9-methoxycarbonyl (Fmoc), cyclopentyloxycarbonyl,adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and thelike; aralkyl groups such as benzyl, triphenylmethyl, benzyloxymethyland the like; and silyl groups such as trimethylsilyl and the like.Additionally, the “amine protecting group” can be a divalent protectinggroup such that both hydrogen atoms on a primary amine are replaced witha single protecting group. In such a situation the amine protectinggroup can be phthalimide (phth) or a substituted derivative thereofwherein the term “substituted” is as defined above. In some embodiments,the halogenated phthalimide derivative may be tetrachlorophthalimide(TCphth).

A “hydroxyl protecting group” is well understood in the art. A hydroxylprotecting group is a group which prevents the reactivity of thehydroxyl group during a reaction which modifies some other portion ofthe molecule and can be easily removed to generate the desired hydroxyl.Hydroxyl protecting groups can be found at least in Greene and Wuts,1999, which is incorporated herein by reference. Some non-limitingexamples of hydroxyl protecting groups include acyl groups such asformyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl,2-bromoacetyl, trifluoroacetyl, trichloroacetyl, o-nitrophenoxyacetyl,α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl,4-nitrobenzoyl, and the like; sulfonyl groups such as benzenesulfonyl,p-toluenesulfonyl and the like; acyloxy groups such as benzyloxycarbonyl(Cbz), p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl,p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl,p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl,3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl,3,4,5-trimethoxybenzyloxycarbonyl,1-(p-biphenylyl)-1-methylethoxycarbonyl,α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl,t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl,isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl(Alloc), 2,2,2-trichloroethoxycarbonyl, 2-trimethylsilylethyloxycarbonyl(Teoc), phenoxycarbonyl, 4-nitrophenoxycarbonyl,fluorenyl-9-methoxycarbonyl (Fmoc), cyclopentyloxycarbonyl,adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and thelike; aralkyl groups such as benzyl, triphenylmethyl, benzyloxymethyland the like; and silyl groups such as trimethylsilyl and the like.

A “thiol protecting group” is well understood in the art. A thiolprotecting group is a group which prevents the reactivity of themercapto group during a reaction which modifies some other portion ofthe molecule and can be easily removed to generate the desired mercaptogroup. Thiol protecting groups can be found at least in Greene and Wuts,1999, which is incorporated herein by reference. Some non-limitingexamples of thiol protecting groups include acyl groups such as formyl,acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl,2-bromoacetyl, trifluoroacetyl, trichloroacetyl, o-nitrophenoxyacetyl,α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl,4-nitrobenzoyl, and the like; sulfonyl groups such as benzenesulfonyl,p-toluenesulfonyl and the like; acyloxy groups such as benzyloxycarbonyl(Cbz), p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl,p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl,p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl,3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl,3,4,5-trimethoxybenzyloxycarbonyl,1-(p-biphenylyl)-1-methylethoxycarbonyl,α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl,t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl,isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl(Alloc), 2,2,2-trichloroethoxycarbonyl, 2-trimethylsilylethyloxycarbonyl(Teoc), phenoxycarbonyl, 4-nitrophenoxy carbonyl, fluorenyl-9-methoxycarbonyl (Fmoc), cyclopentyloxycarbonyl, adamantyloxycarbonyl,cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; aralkyl groupssuch as benzyl, triphenylmethyl, benzyloxymethyl and the like; and silylgroups such as trimethylsilyl and the like.

A “stereoisomer” or “optical isomer” is an isomer of a given compound inwhich the same atoms are bonded to the same other atoms, but where theconfiguration of those atoms in three dimensions differs. “Enantiomers”are stereoisomers of a given compound that are mirror images of eachother, like left and right hands. “Diastereomers” are stereoisomers of agiven compound that are not enantiomers. Chiral molecules contain achiral center, also referred to as a stereocenter or stereogenic center,which is any point, though not necessarily an atom, in a moleculebearing groups such that an interchanging of any two groups leads to astereoisomer. In organic compounds, the chiral center is typically acarbon, phosphorus or sulfur atom, though it is also possible for otheratoms to be stereocenters in organic and inorganic compounds. A moleculecan have multiple stereocenters, giving it many stereoisomers. Incompounds whose stereoisomerism is due to tetrahedral stereogeniccenters (e.g., tetrahedral carbon), the total number of hypotheticallypossible stereoisomers will not exceed 2^(n), where n is the number oftetrahedral stereocenters. Molecules with symmetry frequently have fewerthan the maximum possible number of stereoisomers. A 50:50 mixture ofenantiomers is referred to as a racemic mixture. Alternatively, amixture of enantiomers can be enantiomerically enriched so that oneenantiomer is present in an amount greater than 50%. Typically,enantiomers and/or diastereomers can be resolved or separated usingtechniques known in the art. It is contemplated that that for anystereocenter or axis of chirality for which stereochemistry has not beendefined, that stereocenter or axis of chirality can be present in its Rform, S form, or as a mixture of the R and S forms, including racemicand non-racemic mixtures. As used herein, the phrase “substantially freefrom other stereoisomers” means that the composition contains ≤15%, morepreferably ≤10%, even more preferably ≤5%, or most preferably ≤1% ofanother stereoisomer(s).

VII. EXAMPLES

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventor to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the spirit and scope ofthe invention.

Example 1—Synthesis of Trioxacarcin and Analogs Thereof

The polyoxygenated 2,7-dioxabicyclo[2.2.1]heptane system of thetrioxacarcins is a most intriguing structural motif requiring specialattention with regard to strategy and experimentation for itsconstruction. FIG. 2 presents, in one embodiment, a designed strategytoward DC-45-A2 (1) in retrosynthetic format. Thus, disconnection of thehemiacetal moiety of 1 followed by functional group transformations ledto advanced precursor 5, whose conversion to the target molecule couldbe envisioned through sequential and selective deprotection/oxidations.Dismantling of the bicyclo[2.2.1]heptane system within 5 through anepoxyketone rearrangement (Gaoni, 1968; Waserman et al., 1969; Wassermanet al., 1986a; Wasserman et al., 1986b; Wasserman et al., 1988a;Wasserman et al., 1988b; Naruse et al., 1988a; Naruse et al., 1988b;Evans et al., 1991) revealed epoxyketone 6 as a precursor, whose origincould be traced back to key building blocks 7-10 through thedisconnections indicated in FIG. 2 [e.g. a) Hauser-Kraus annulation; b)Stille reaction; c) asymmetric Jorgensen epoxidation; and d)Baylis-Hillman reaction]. The key epoxyketone rearrangement (6→5, FIG.2) was presumed to be inducible in a stereo- and regioselective mannerthrough the action of a suitable monodentate Lewis acid that wouldinvolve inversion of configuration at C6, as indicated in FIG. 2 (seearrows on structure 6).

The required cyclohexenone 10 was prepared enantioselectively fromcyclohexadiene 11, as summarized in Scheme 1. Thus, 11 was subjected toUpjohn dihydroxylation (OsO₄ cat., NMO, 50% yield) and the resultingdiol 12 was silylated to afford bis-TBS ether 13 (TBSCl, 92% yield).Epoxidation of the latter (mCPBA, 89% yield) led selectively to epoxide14, whose regioselective opening with (−)-norephedrine-derived amine 15in the presence of nBuLi furnished allylic alcohol 16 in 94% yield and89% ee (Maras et al., 1998; O'Brien et al., 1998; Coleman et al., 1999;deSousa et al., 2002). Protection of this alcohol with4-methoxybenzyl-2,2,2-trichloroacetimidate (PMBTCA) followed byTBAF-induced desilylation led to PMB-ether diol 17 in 84% yield.Selective oxidation of the allylic alcohol of the latter (TEMPO, pTSA,74% yield) (Banwell et al., 1994) furnished hydroxyenone 18, whosesilylation (TBSCl, 99% yield) led to the targeted key building blockenone 10.

Enone 10 was coupled with the easily accessible iodocyanophthalidederivative 9 through a Hauser-Kraus annulation, (Hauser et al., 1978;Kraus et al., 1978) and the product was elaborated to intermediate 21 asshown in Scheme 2. Thus, iodocyanophthalide 9 [prepared from the knowncyanophthalide 19 (Nicolaou et al., 2009) by sequential iodination (NIS)and MOM protection (MOMCl, DIPEA, 50% overall yield)] was reacted withenone 10 in the presence of tBuOLi (−78° C.) (S̆venda et al., 2011;Magauer et al., 2013; Nicolaou et al., 2009) and the resultingp-dihydroquinone derivative was selectively methylated with Me₂SO₄ toafford tricyclic system 20 in 69% overall yield. Removal of the MOMgroup from the latter intermediate with MgBr₂.OEt₂, (Yang et al., 2009)followed by treatment with tBu₂Si(OTf)₂ and 2,6-lutidine then gavesilylated product 21 in 85% overall yield.

Intermediate 21 was advanced to the key cyclization precursor 6, assummarized in Scheme 3. Thus, Stille coupling of aryl iodide 21 withstannane 8 (Pilli et al., 1998) proceeded in the presence of CuTC andcatalytic amounts of Pd(PPh₃)₄ (Pulukuri et al., 2012) to afford allylicalcohol 22 (74% yield), whose oxidation with TEMPO and PIDA gavealdehyde 23 (89% yield). Jorgensen asymmetric epoxidation ofα,β-unsaturated aldehyde 23 (24 cat., urea.H₂O₂) (Marigo et al., 2005)led to epoxyaldehyde 25, which was subjected without purification toBaylis-Hillman reaction with enone 7 (Edwards et al., 2003) (DABCO,4-nitrophenol) to give labile hydroxyepoxide 26. The latter wasimmediately protected with N-trimethylsilylimidazole (TMS-imid) tofurnish the targeted precursor 6 (+C4-epi-6, d.r. ca. 3:1) in 36% yieldover the three steps.

With the penultimate bis-cyclization precursor 6 in hand, the stage wasnow set for the coveted cascade ring closures to forge the targeted2,7-dioxabicyclo[2.2.1]heptane system of the growing molecule. To thisend, and as shown in Scheme 4, epoxyketone 6 (ca. 3:1 mixture ofC4-diastereoisomers) was reacted with catalytic amounts of BF₃.OEt₂(monodentate Lewis acid) in CH₂Cl₂ at −78° C., furnishing the desiredproduct as a mixture of C4-diastereoisomers (d.r. ca. 3:1) (5a,C4-α-diastereoisomer, 54% yield; 5b, C4-β-diastereoisomer, 18% yield).The assignments of the C4 and C6 configurations of diastereoisomers 5aand 5b were based on their H4, H5, H6 coupling constants (5a:J_(4,5)=4.8 Hz, J_(5,6)=3.0 Hz; 5b: J_(4,5)=0 Hz, J_(5,6)=3.6 Hz).(Padwa et al., 1991; Kraehenbuehl et al., 1995; Kraehenbuehl et al.,1998; Muthusamy et al., 2002). Both compounds were obtained as singlediastereoisomers at C6 (inverted configuration). The reaction ispresumed to proceed through transition states TS-6⋅B and TS′-6⋅B asshown in Scheme 4. In contrast, reaction of substrate 6 with catalyticamounts of bidentate Lewis acid SnCl₄ in CH₂Cl₂ at −78° C. led to theopposite diastereoisomers at C6 (J_(5,6)=0 Hz), 27 (+C4-epi-27) (37%yield, d.r. ca. 13:1). This reaction is presumed to proceed throughtransition states TS-6⋅Sn and TS′-6⋅Sn, the latter being favored overits more sterically congested alternative conformer TS″-6⋅Sn that wouldhave led to inversion of configuration at C6 (see Scheme 4). Theseresults support the proposed monodentate Lewis acid-catalyzedepoxyketone rearrangement upon which the strategy for the constructionof the dioxabicyclo[2.2.1]heptane structural motif possessing thedesired configurations was based.

Having succeeded in building the most challenging structural domain ofthe targeted molecule, the remaining tasks of the synthesis werecompleted including installation of the epoxide moiety, oxidation at C4,and deprotection. Thus, advanced intermediate 5a (major diastereoisomer)was converted to the targeted natural product (1) as shown in Scheme 5.Thus, selective cleavage of the TMS-ether of 5a gave allylic alcohol 28(TFA, 65%) and recovered 5a (24% yield). Due to difficulties inobtaining the desired epoxide 29 from 28 through mCPBA ortBuOOH/VO(acac)₂ epoxidations, a three-step process involvingdiastereoselective Upjohn dihydroxylation of the olefinic bond within 28(OsO₄ cat., NMO) was used followed by selective monotosylation of theresulting triol (TsCl, Et₃N, DMAP cat.) and epoxide formation (K₂CO₃,MeOH, 82% overall yield). TPAP-catalyzed oxidation of hydroxyepoxide 29led to ketoepoxide 30 (93% yield), which could be sequentially andselectively deprotected to afford trioxacarcin derivatives 31 (Et₃N.3HF,3.0 equiv, 15 min, 88% yield) and 32 (DDQ, 93% yield). Finally,trioxacarcin DC-45-A2 (1) was liberated from its TBS-ether 32 byexposure to Et₃N.3HF (excess, 13 h, 86% yield). Synthetic 1 exhibitedidentical physical properties (i.e., ¹H and ¹³C NMR and mass spectra) tothose reported in the literature (Shirahata et al., 1984; S̆venda et al.,2011).

Example 2—General Methods and Materials

All reactions were carried out under an argon atmosphere with drysolvent under anhydrous conditions, unless otherwise noted. Dryacetonitrile (CH₃CN), N,N-dimethylformamide (DMF), methanol,dichloromethane (CH₂Cl₂), triethylamine (Et₃N) and tetrahydrofuran (THF)were obtained by passing commercially available pre-dried, oxygen-freeformulations through activated alumina columns. Anhydrous acetone,cyclohexane, chloroform (CHCl₃) and 1,2-dichloroethane (DCE) werepurchased from commercial suppliers and stored under argon. Yields referto chromatographically and spectroscopically (¹H NMR) homogenousmaterial, unless otherwise stated. Reagents were purchased at thehighest commercial quality and used without further purification, unlessotherwise noted. Reactions were monitored by thin-layer chromatography(TLC) carried out on 0.25 mm E. Merck silica gel plates (60F₂₅₄) usingUV light as visualizing agent or an aqueous solution of phosphomolybdicacid and cerium sulfate or a basic aqueous solution of potassiumpermanganate and heat as developing agents. Acros Organics silica gel(60, particle size 0.035-0.070 mm) was used for flash columnchromatography.

NMR spectra were recorded on a Bruker Avance III HD 600 MHz instrumentequipped with a 5 mm DCH cryoprobe and calibrated using residualundeuterated solvent (CDCl₃, δ_(H)=7.26 ppm, δ_(C)=77.00 ppm) as aninternal reference at 298 K. The following abbreviations were used todesignate multiplicities: s=singlet, d=doublet, t=triplet, q=quartet,m=multiplet, br=broad. Infrared (IR) spectra were recorded on aPerkin-Elmer 100 FT-IR spectrometer. High-resolution mass spectra (HRMS)were recorded on an Ion Trap-Time of Flight Mass Spectrometer (Shimadzu,Columbia, Md.) operated with an ESI source interface and a VG ZAB-ZSEmass spectrometer using MALDI (matrix-assisted laser-desorptionionization) or ESI (electrospray ionization). Optical rotations wererecorded on a Schmidt+Haensch Polartronic M100 polarimeter at 589.44 nmusing 100 mm cells and the solvent and concentration indicated.

Example 3—Compound Characterization

Diol 12:

To a stirred solution of N-methylmorpholine-N-oxide (42.2 g, 312 mmol)in acetone (780 mL) at 25° C. were added 1,4-cyclohexadiene (25.0 g, 312mmol, 1.0 equiv) and OsO₄ (4% w/v aq. solution, 39.7 mL, 6.24 mmol, 0.02equiv), and the resulting black suspension was stirred at thistemperature for 72 h. Na₂SO₃ (25.0 g) was then added and the resultingmixture was stirred at 25° C. for another 1 h. The mixture was driedover anhydrous MgSO₄ and concentrated under reduced pressure. Theresidue was filtered through a short pad of silica gel, rinsed withEtOAc and concentrated to give the title compound (12, 17.9 g, 157 mmol,50%) as a colorless solid. 12: R_(f)=0.23 (silica gel, EtOAc); m.p.71-72° C. (EtOAc); FT-IR (neat): ν_(max)=3292, 3022, 2906, 1649, 1433,1371, 1331, 1079, 1057, 1049, 975, 895, 755, 662 cm⁻¹; ¹H NMR (CDCl₃,600 MHz) δ=5.58 (t, J=1.8 Hz, 2H), 3.94 (t, J=6.0 Hz, 2H), 2.36 (dd,J=6.0, 16.2 Hz, 2H), 2.25 (dd, J=16.2, 6.0 Hz, 2H), 2.20 (br s, 1H),2.17 (br s, 1H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=123.7, 68.9, 31.0 ppm;HRMS (ESI-TOF) calcd for C₆H₁₀NaO₂ ⁺ [M+Na]⁺ 137.0573, found 137.0572.All spectroscopic data were consistent with those in the literature.(Mara et al., 1998)

Bis-TBS Ether 13:

To a stirred solution of diol 12 (6.27 g, 54.9 mmol) in CH₂Cl₂ at 25° C.were added imidazole (18.7 g, 274 mmol, 5.0 equiv) and TBSCl (19.9 g,132 mmol, 2.4 equiv). After stirring at this temperature for 48 h, thereaction mixture was washed with water (2×100 mL), whereupon thecombined aqueous layers were extracted with CH₂Cl₂ (5×100 mL). Thecombined organic phases were dried over anhydrous MgSO₄ and concentratedunder reduced pressure. The crude mixture was purified by flash columnchromatography (silica gel, EtOAc:hexanes 1:200) to give the titlecompound (13, 17.4 g, 50.7 mmol, 92%) as a colorless oil. 13: R_(f)=0.37(silica gel, EtOAc:hexanes 1:100); FT-IR (neat): ν_(max)=2954, 2927,2894, 2857, 1472, 1373, 1250, 1215, 1121, 1094, 1074, 1006, 827, 772cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=5.50 (t, J=1.8 Hz, 2H), 3.84 (t, J=5.4Hz, 2H), 2.21 (dd, J=16.2, 6.6 Hz, 2H), 2.13 (dd, J=16.2, 5.4 Hz, 2H),0.88 (s, 18H), 0.06 (s, 6H), 0.05 (s, 6H) ppm; NMR (CDCl₃, 150 MHz)δ=124.1, 70.7, 32.6, 25.9, 18.2, −4.4, −4.9 ppm; HRMS (ESI-TOF) calcdfor C₁₈H₃₉O₂Si₂ ⁺ [M+H]⁺ 343.2483, found 343.2474. All spectroscopicdata were consistent with those in the literature. (O'Brien et al.,1998)

Epoxide 14:

To a stirred solution of bis-TBS ether 13 (16.7 g, 48.8 mmol) incyclohexane (500 mL) at 25° C. were sequentially added NaHCO₃ (8.19 g,97.5 mmol, 2.0 equiv) and mCPBA (11.8 g, ca. 30% water content, 68.3mmol, 1.4 equiv) in portions. The reaction mixture was stirred at thistemperature for 17 h. After quenching the reaction with Na₂SO₃ (10% aq.,300 mL), the aqueous layer was extracted with CH₂Cl₂ (2×400 mL), and thecombined organic phases were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography (silica gel, EtOAc:hexanes 1:30) to give the titlecompound (14, 15.5 g, 43.2 mmol, 89%) as a colorless oil. 14: R_(f)=0.32(silica gel, EtOAc:hexanes 1:30); FT-IR (neat): ν_(max)=2952, 2928,2894, 2856, 1472, 1371, 1250, 1135, 1101, 1076, 998, 956, 879, 827, 805,773 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=3.67 (t, J=4.8 Hz, 2H), 3.15 (s,2H), 2.08-2.00 (m, 4H), 0.88 (s, 18H), 0.05 (s, 6H), 0.04 (s, 6H) ppm;¹³C NMR (CDCl₃, 150 MHz) δ=68.8, 52.1, 31.3, 25.9, 18.2, −4.5, −4.8 ppm;HRMS (ESI-TOF) calcd for C₁₈H₃₉O₃Si₂ ⁺ [M+H]⁺ 359.2427, found 359.2432.All spectroscopic data were consistent with those in the literature.(O'Brien et al., 1998)

(−)-Norephedrine-Derived Amine 15:

Amine 15 was prepared according to the original procedure. (Colman etal., 1999) 15: R_(f)=0.10 (silica gel, 10% MeOH in CH₂Cl₂); [α]²⁵_(D)=−22.8 (c=1.0, CHCl₃); ¹H NMR (CDCl₃, 600 MHz) δ=7.36-7.32 (m, 4H),7.25-7.21 (m, 1H), 3.85 (d, J=3.0 Hz, 1H), 2.64-2.62 (m, 4H), 2.34 (s,3H), 2.27 (qd, J=6.6, 3.6 Hz, 1H), 1.82-1.80 (m, 4H), 1.70 (br s, 1H),0.85 (d, J=6.6 Hz, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=141.9, 128.0,127.9, 126.5, 67.0, 66.3, 52.4, 35.5, 23.5, 13.0 ppm. All spectroscopicdata were consistent with those in the literature. (Colman et al., 1999)

Allylic Alcohol 16:

To a stirred solution of (−)-norephedrine-derived amine 15 (4.4 g, 20.2mmol, 2.0 equiv) in THF (14 mL) at −78° C. was added nBuLi (2.1 M inhexanes, 9.6 mL, 20.2 mmol, 2.0 equiv) dropwise over 20 min. Afterwarming the reaction mixture to 0° C. and stirring at this temperaturefor 30 min, the reaction was cooled to −78° C. again, and a solution ofepoxide 14 (3.6 g, 10.1 mmol) in THF (14 mL) was added dropwise at −78°C. over 20 min. The reaction mixture was allowed to warm to 25° C.,stirred at this temperature for 18 h, and then quenched with NH₄Cl (sat.aq., 25 mL). The resulting mixture was extracted with Et₂O (2×40 mL) andthe combined organic phases were washed sequentially with HCl (2% aq.,3×40 mL), NaHCO₃ (sat. aq., 2×40 mL) and brine (25 mL), and then driedover anhydrous Na₂SO₄ and concentrated under reduced pressure. Theresidue was purified by flash column chromatography (silica gel,EtOAc:hexanes 1:5) to give the title compound (16, 3.4 g, 9.5 mmol, 94%,19:1 e.r. by Mosher ester analysis—Hoye et al., 2007) as a white solid.16: R_(f)=0.34 (silica gel, EtOAc:hexanes 1:4); m.p. 56-57° C. (EtOAc,hexanes); [α]²⁵ _(D)=−96.3=1.0, CHCl₃); FT-IR (neat): ν_(max)=3302,2953, 2929, 2886, 2856, 1472, 1389, 1251, 1118, 1091, 1030, 953, 871,829, 772, 672 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=5.74 (dd, J=10.1, 2.6 Hz,1H), 5.64 (dd, J=10.1, 3.1 Hz, 1H), 4.45-4.43 (m, 1H), 4.08-4.06 (m,1H), 2.27 (ddd, J=13.3, 8.1, 5.3 Hz, 1H), 1.57 (ddd, J=13.2, 6.3, 2.2Hz, 1H), 1.46 (d, J=3.8 Hz, 1H), 0.90 (s, 9H), 0.89 (s, 9H), 0.08 (s,3H), 0.08 (s, 3 H), 0.08 (s, 3H), 0.07 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150MHz) δ=131.1, 130.3, 69.7, 68.8, 65.9, 37.4, 26.0, 25.9, 18.4, 18.3,−4.4, −4.6, −4.8 ppm; HRMS (ESI-TOF) calcd for C₁₈H₃₈NaO₃Si₂ ⁺ [M+Na]⁺381.2252, found 381.2235. All spectroscopic data were consistent withthose in the literature. (de Sousa et al., 2002)

PMB-Ether Diol 17:

To a stirred solution of allylic alcohol 16 (17.0 g, 47.4 mmol) in THF(500 mL) 25° C. were added trityltetrafluoroborate (0.782 g, 2.37 mmol,0.05 equiv) and freshly prepared4-methoxybenzyl-2,2,2-trichloroacetimidate (PMB-TCA; 33.5 g, 118 mmol,2.5 equiv). After stirring at this temperature for 1 h, TBAF (1.0 M inTHF, 332 mL, 7.0 equiv) was added and the reaction mixture was heated toreflux for 4 h, then cooled to 25° C. and concentrated under reducedpressure. The residue was purified by flash column chromatography(silica gel, acetone:pentane 1:3→3:2) to give the title compound (17,10.0 g, 40.0 mmol, 84%) as an off-white solid. 17: R_(f)=0.38 (silicagel, acetone:pentane 2:3); m.p. 84-85° C. (acetone, pentane); [α]²⁵_(D)=−135.3 (c=1.0, CHCl₃); FT-IR (neat): ν_(max)=3373, 3031, 2931,2837, 1611, 1512, 1388, 1301, 1244, 1172, 1063, 1031, 823 cm⁻¹; ¹H NMR(CDCl₃, 600 MHz) δ=7.26 (d, J=8.5 Hz, 2H), 6.87 (d, J=8.5 Hz, 2H), 5.96(dd, J=10.1, 2.3 Hz, 1H), 5.76 (dd, J=10.1, 2.7 Hz, 1H), 4.52 (d, J=11.3Hz, 1H), 4.48 (d, J=11.3 Hz, 1H), 4.17-4.13 (m, 3H), 3.79 (s, 3H), 2.50(s, 1H), 2.18 (ddd, J=13.1, 8.1, 4.9 Hz, 1H), 1.81 (ddd, J=13.4, 6.2,2.4 Hz, 1H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=159.2, 130.6, 130.4, 129.4,129.3, 113.8, 70.9, 70.4, 67.4, 66.7, 55.3, 32.7 ppm; HRMS (ESI-TOF)calcd for C₁₄H₁₈NaO₄ ⁺ [M+Na]⁺ 273.1097, found 273.1086.

Hydroxyenone 18:

To a stirred solution of PMB-ether diol 17 (6.09 g, 24.3 mmol) in CH₂Cl₂(290 mL) at 25° C. was added p-toluene sulfonic acid monohydrate (13.9g, 72.9 mmol, 3.0 equiv). Then a solution of TEMPO (11.4 g, 72.9 mmol,3.0 equiv) in CH₂Cl₂ (30 mL) was added via syringe pump over 30 min at0° C. After stirring at this temperature for another 15 min, thereaction was quenched with NaHCO₃ (sat. aq., 150 mL). The resultingmixture was extracted with CH₂Cl₂ (2×100 mL), and the combined organicphases were washed with brine (200 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography (silica gel, EtOAc:hexanes 1:3→4:1) to give thetitle compound (18, 4.47 g, 18.0 mmol, 74%) as an orange oil. 18:R_(f)=0.40 (silica gel, EtOAc:hexanes 1:1); [α]²⁵ _(D)=−158.8 (c=1.0,CHCl₃); FT-IR (neat): ν_(max)=3449, 2934, 2865, 2838, 1691, 1612, 1513,1247, 1173, 1106, 1060, 1033, 832 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=7.27(d, J=8.4 Hz, 2H), 6.89-6.86 (m, 3H), 6.07 (d, J=10.0 Hz, 1H), 4.64-4.59(m, 2H), 4.51 (d, J=11.3 Hz, 1H), 4.23-4.21 (m, 1H), 3.79 (s, 3H), 3.53(s, 1H), 2.64 (dt, J=13.2, 2.4 Hz, 1H), 1.95 (dt, J=13.2, 4.2 Hz, 1H)ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=200.3, 159.4, 146.6, 129.6, 129.3,127.7, 113.9, 71.5, 69.7, 68.9, 55.2, 35.0 ppm; HRMS (ESI-TOF) calcd forC₁₄H₁₇O₄ ⁺ [M+H]⁺ 249.1121, found 249.1114. All spectroscopic data wereconsistent with those in the literature. (Kato et al., 2006; Myers etal., 2011)

Enone 10:

To a stirred solution of hydroxyenone 18 (4.47 g, 18.0 mmol) in CH₂Cl₂(90 mL) at 25° C. were added imidazole (3.67 g, 54.0 mmol, 3.0 equiv)and TBSCl (4.88 g, 32.4 mmol, 1.8 equiv). After stirring at thistemperature for 1.5 h, the reaction was quenched with NH₄Cl (sat. aq.,100 mL). The resulting mixture was extracted with CH₂Cl₂ (2×50 mL), andthe combined organic phases were washed with brine (200 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The residuewas purified by flash column chromatography (silica gel, EtOAc:hexanes1:3) to give the title compound (10, 6.45 g, 17.8 mmol, 99%) as acolorless oil. 10: R_(f)=0.54 (silica gel, EtOAc:hexanes 1:4); [α]²⁵_(D)=−119.5 (c=1.0, CHCl₃); FT-IR (neat): ν_(max)=2953, 2929, 2885,2856, 1696, 1612, 1513, 1249, 1172, 1147, 1077, 1036, 835, 779 cm⁻¹; ¹HNMR (CDCl₃, 600 MHz) δ=7.28 (d, J=8.5 Hz, 2H), 6.90 (d, J=8.6 Hz, 2H),6.87 (dd, J=10.3, 3.6 Hz, 1H), 5.95 (d, J=10.2 Hz, 1H), 4.60 (d, J=11.4Hz, 1H), 4.55 (d, J=11.4 Hz, 1H), 4.39-4.35 (m, 2H), 3.81 (s, 3H),2.30-2.20 (m, 2H), 0.88 (s, 9H), 0.10 (s, 3H), 0.09 (s, 3H) ppm; ¹³C NMR(CDCl₃, 150 MHz) δ=197.4, 159.5, 147.8, 129.8, 129.5, 128.2, 114.0,71.2, 71.1, 70.3, 55.3, 37.7, 25.7, 18.3, 4.7, −5.4 ppm; HRMS (ESI-TOF)calcd for C₂₀H₃₀NaO₄Si⁺ [M+Na]⁺ 385.1806, found 385.1798. Allspectroscopic data were consistent with those in the literature. (Katoet al., 2006; Myers et al., 2011)

Cyanophthalide 19:

The cyanophthalide 19 was prepared according to the reported procedure.(Nicolaou et al., 2009) 19: R_(f)=0.52 (silica gel, EtOAc:hexanes 1:1);¹H NMR (CDCl₃, 600 MHz) δ=7.40 (s, 1H), 6.99 (s, 1H), 6.91 (s, 1H), 6.00(s, 7H), 2.49 (s, 4H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=168.9, 156.5,150.9, 141.9, 118.6, 115.0, 113.6, 107.0, 66.0, 22.5 ppm. Allspectroscopic data were consistent with those reported in theliterature. (Nicolaou et al., 2009)

Iodocyanophthalide 9:

To a stirred solution of cyanophthalide 19 (1.04 g, 5.46 mmol) in DCE(100 mL) at −10° C. was added NIS (1.61 g, 7.11 mmol, 1.3 equiv). Thereaction flask was covered with aluminum foil to exclude light. Afterstirring at this temperature for 6 h, the reaction was quenched withNa₂SO₃ (10% aq., 100 mL). The resulting mixture was extracted withCH₂Cl₂ (2×50 mL), and the combined organic phases were dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The residuewas subjected to flash column chromatography (silica gel, EtOAc:hexanes1:1) to give the unprotected iodocyanophthalide intermediate as amixture with some starting material 19 (ratio of 10:1).

To a solution of the above mixture in CH₂Cl₂ (50 mL) at 25° C. wereadded chloromethyl methyl ether (376 mg, 4.64 mmol, 0.85 equiv) andN,N-diisopropylethylamine (478 mg, 3.71 mmol, 0.68 equiv). Afterstirring at this temperature for 6 h, the reaction was quenched withNaHCO₃ (5% aq., 100 mL). The resulting mixture was extracted with CH₂Cl₂(2×50 mL), and the combined organic phases were dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The residue was purifiedby flash column chromatography (silica gel, EtOAc: hexanes 1:6) to givethe title compound (9, 984 mg, 2.73 mmol, 50%, two steps) as a yellowsolid. 9: R_(f)=0.33 (silica gel, EtOAc:hexanes 1:4); m.p. 117-118° C.(EtOAc: hexanes); FT-IR (neat): ν_(max)=2930, 1780, 1599, 1452, 1379,1262, 1206, 1156, 1034, 983, 890, 770 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz)δ=7.29 (s, 1H), 5.93 (s, 1H), 5.54 (d, J=6.2 Hz, 1H), 5.50 (d, J=6.2 Hz,1H), 3.68 (s, 3H), 2.67 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ 164.3,156.4, 153.3, 143.4, 118.1, 113.5, 112.2, 102.5, 101.6, 64.7, 59.0, 30.5ppm; HRMS (ESI-TOF) calcd for C₁₂H₉IO₄Si⁻ [M−H]⁻ 357.9582, found357.9575.

Aryl Iodide 20:

To a stirred solution of iodocyanophthalide 9 (889 mg, 2.46 mmol) in THF(27 mL) at −78° C. was added tBuOLi (1.0 M in THF, 7.40 mL, 0.740 mmol,3.0 equiv). After stirring at this temperature for 10 min, a solution ofenone 10 (889 mg, 2.46 mmol, 1.0 equiv) in THF (27 mL) was addeddropwise. The resulting reaction mixture was stirred at −78° C. for 30min before Me₂SO₄ (3.05 g, 24.6 mmol, 10 equiv) was added dropwise. Theresulting mixture was warmed to −5° C. and stirred at this temperaturefor 5 h before it was quenched with NH₄Cl (sat. aq., 150 mL). Theresulting mixture was extracted with EtOAc (3×80 mL), and the combinedorganic phases were washed with brine (150 mL), dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The residue was purifiedby flash column chromatography (silica gel, EtOAc:hexanes 1:50) to givethe title compound 20 (1.24 g, 1.70 mmol, 69%) as an orange oil. 20:R_(f)=0.62 (silica gel, EtOAc:hexanes 1:8); [α]²⁵ _(D)=+37.0 (c=1.0,CHCl₃); FT-IR (neat): ν_(max)=2952, 2929, 2855, 1635, 1611, 1514, 1441,1361, 1250, 1158, 1043, 1003, 924, 872, 837, 780 cm⁻¹; ¹H NMR (CDCl₃,600 MHz) δ=14.87 (s, 1H), 7.73 (s, 1H), 7.28 (d, J=8.6 Hz, 2H), 6.87 (d,J=8.6 Hz, 2H), 5.21 (d, J=5.6 Hz, 1H), 5.18 (t, J=2.6 Hz, 1H), 5.15 (d,J=5.6 Hz, 1H), 4.99 (dd, J=12.4, 5.2 Hz, 1H), 4.68 (d, J=11.1 Hz, 1H),4.58 (d, J=11.0 Hz, 1H), 3.85 (s, 3H), 3.79 (s, 3H), 3.78 (s, 3H), 2.72(ddd, J=13.4, 5.1, 3.4 Hz, 1H), 2.67 (s, 3H), 2.18 (ddd, J=13.3, 2.3 Hz,1H), 1.55 (s, 3H), 0.98 (s, 9H), 0.25 (s, 3H), 0.18 (s, 3H) ppm; ¹³C NMR(CDCl₃, 150 MHz) δ 204.0, 160.1, 159.3, 156.0, 145.3, 144.8, 134.3,130.1, 129.5, 126.6, 118.5, 118.4, 113.9, 108.9, 101.7, 100.3, 70.9,69.4, 68.7, 62.9, 59.1, 55.3, 36.5, 30.3, 25.9, 18.5, −4.4, −5.3 ppm;HRMS (ESI-TOF) calcd for C₃₂H₄₁INaO₈Si⁺ [M+Na]⁺ 731.1508, found731.1485.

Aryl Iodide 21:

To a stirred solution of aryl iodide 20 (1.24 g, 1.70 mmol) in THF (35mL) at 0° C. was added MgBr₂.OEt₂ (1.32 g, 5.10 mmol, 3.0 equiv) in oneportion. After stirring at this temperature for 10 min, the reaction wasquenched with H₂O (50 mL). The resulting mixture was extracted withEtOAc (3×50 mL), and the combined organic phases were dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The cruderesidue was taken to the next step without further purification. To astirred solution of the above crude in DMF (17 mL) at 0° C. was added2,6-lutidine (546 mg, 5.10 mmol, 3.0 equiv), and then tBu₂Si(OTf)₂ (896mg, 2.04 mmol, 1.2 equiv) was added dropwise over a period of 10 min.After stirring at this temperature for another 10 min, the reaction wasquenched with NH₄Cl (sat. aq., 50 mL) and diluted with EtOAc (100 mL).The resulting mixture was washed with brine (3×100 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The residuewas purified by flash column chromatography (silica gel, EtOAc: hexanes1:50) to give the title compound (21, 1.16 g, 1.45 mmol, 85%, two steps)as an orange oil. 21: R_(f)=0.54 (silica gel, EtOAc:hexanes 1:4); [α]²⁵_(D)=+31.7 (c=1.0, CHCl₃); FT-IR (neat): ν_(max)=2933, 2896, 2859, 1701,1600, 1560, 1514, 1471, 1399, 1359, 1249, 1157, 1064, 1007, 828, 780cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=7.50 (s, 1H), 7.31 (d, J=8.6 Hz, 2H),6.87 (d, J=8.6 Hz, 2H), 5.19 (t, J=2.8 Hz, 1H), 4.87 (dd, J=12.4, 5.0Hz, 1H), 4.72 (d, J=10.8 Hz, 1H), 4.62 (d, J=10.8 Hz, 1H), 3.89 (s, 3H),3.79 (s, 3H), 2.73 (ddd, J=13.6, 5.0, 3.1 Hz, 1H), 2.62 (s, 3H), 2.15(dt, J=13.2, 2.4 Hz, 1H), 1.55 (s, 3H), 1.14 (s, 9H), 1.11 (s, 9H), 0.96(s, 9H), 0.24 (s, 3H), 0.14 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz)δ=194.4, 159.3, 152.3, 148.9, 146.3, 143.1, 131.2, 130.2, 129.8, 129.6,115.3, 114.7, 114.6, 113.9, 90.7, 71.3, 71.2, 69.9, 62.7, 55.3, 36.4,29.6, 26.1, 26.0, 26.0, 21.3, 20.9, 18.7, −4.3, −5.4 ppm; HRMS (ESI-TOF)calcd for C₃₈H₅₄IO₇Si₂ ⁺ [M+H]⁺ 805.2447, found 805.2470.

Stannane 8:

The stannane 8 was prepared according to the original procedure. (Philliet al., 1998) 8: R_(f)=0.51 (silica gel, EtOAc:hexanes 1:8); ¹H NMR(CDCl₃, 600 MHz) δ 6.29-6.21 (m, 1H), 4.17 (s, 1H), 1.52-1.47 (m, 6H),1.34-1.27 (m, 6H), 0.94-0.84 (m, 15H) ppm; ¹³C NMR (CDCl₃, 150 MHz)δ=147.0, 128.3, 66.4, 29.0, 27.3, 13.7, 9.4 ppm. All spectroscopic datawere consistent with those reported in the literature. (Pilli et al.,1998)

Allylic Alcohol 22:

To a stirred mixture of aryl iodide 21 (1.06 g, 1.32 mmol), CuTC (302mg, 1.58 mmol, 1.2 equiv) and Pd(PPh₃)₄ (305 mg, 0.264 mmol, 0.2 equiv)in DMF (26 mL) was added stannane 8 (642 mg, 1.85 mmol, 1.4 equiv).After stirring at 110° C. for 12 h, the reaction was cooled to 25° C.,then quenched with water (40 mL) and diluted with EtOAc (50 mL). Theresulting mixture was washed with brine (3×50 mL), dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The residue was purifiedby flash column chromatography (silica gel, EtOAc:hexanes 1:4) to givethe title compound (22, 718 mg, 0.977 mmol, 74%) as a yellow foam. 22:R_(f)=0.23 (silica gel, EtOAc:hexanes 1:4); [α]²⁵ _(D)=+26.8 (c=0.85,CH₂Cl₂); FT-IR (neat): ν_(max)=3474, 2896, 2934, 2859, 1697, 1608, 1514,1445, 1371, 1250, 1158, 1124, 1058, 1034, 1010, 938, 880, 829, 781, 662cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=7.40 (s, 1H), 7.32 (d, J=8.6 Hz, 2H),6.87 (d, J=8.6 Hz, 2H), 6.71 (d, J=16.1 Hz, 1H), 6.59 (dt, J=16.1, 5.7Hz, 1H), 5.19 (t, J=2.8 Hz, 1H), 4.88 (dd, J=12.4, 5.0 Hz, 1H), 4.73 (d,J=10.8 Hz, 1H), 4.62 (d, J=10.8 Hz, 1H), 4.39 (dd, J=5.7, 1.3 Hz, 2H),3.90 (s, 3H), 3.79 (s, 3H), 2.73 (ddd, J=13.5, 5.0, 3.1 Hz, 1H), 2.52(s, 3H), 2.15 (dt, J=13.4, 2.9 Hz, 1H), 1.13 (s, 9H), 1.10 (s, 9H), 0.96(s, 9H), 0.25 (s, 3H), 0.14 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz)δ=194.5, 159.3, 150.9, 149.9, 146.2, 134.1, 130.5, 130.3, 129.7, 129.0,124.4, 120.9, 116.1, 115.3, 114.3, 113.9, 71.4, 71.2, 69.9, 64.9, 62.6,55.3, 36.5, 26.2, 26.1, 26.0, 22.2, 21.3, 20.9, 18.7, −4.3, −5.4 ppm;HRMS (ESI-TOF) calcd for C₄₁H₅₉O₈Si₂ ⁺ [M+H]⁺ 735.3743, found 735.3765.

Aldehyde 23:

To a stirred solution of allylic alcohol 22 (600 mg, 0.799 mmol) inCH₂Cl₂ (8 mL) at 25° C. were added TEMPO (12.5 mg, 0.08 mmol, 0.1 equiv)and PhI(OAc)₂ (334 mg, 1.04 mmol, 1.3 equiv). After stirring at thistemperature for 4 h, the reaction was quenched with Na₂SO₃ (10% aq., 20mL). The resulting mixture was extracted with CH₂Cl₂ (3×10 mL), and thecombined organic phases were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography (silica gel, EtOAc:hexanes 1:11) to give the titlecompound (23, 539 mg, 0.720 mmol, 89%) as a yellow foam. 23: R_(f)=0.55(silica gel, EtOAc:hexanes 1:4); [α]²⁵ _(D)=+18.4 (c=1.0, CH₂Cl₂); FT-IR(neat): ν_(max)=2934, 2898, 2859, 1685, 1595, 1514, 1471, 1446, 1372,1249, 1158, 1125, 1060, 1034, 1010, 939, 879, 828, 781, 663 cm⁻¹; ¹H NMR(CDCl₃, 600 MHz) δ=9.72 (d, J=7.7 Hz, 1H), 7.75 (d, J=16.1 Hz, 1H), 7.45(s, 1H), 7.31 (d, J=8.6 Hz, 2H), 7.06 (dd, J=16.1, 7.7 Hz, 1H), 6.87 (d,J=8.6 Hz, 2H), 5.20 (t, J=2.7 Hz, 1H), 4.88 (dd, J=12.4, 5.0 Hz, 1H),4.73 (d, J=10.8 Hz, 1H), 4.63 (d, J=10.8 Hz, 1H), 3.90 (s, 3H), 3.79 (s,3H), 2.74 (ddd, J=13.8, 4.8, 3.0 Hz, 1H), 2.62 (s, 3H), 2.18-2.13 (m,1H), 1.14 (s, 9H), 1.11 (s, 9H), 0.96 (s, 9H), 0.24 (s, 3H), 0.14 (s,3H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=195.3, 194.3, 159.4, 154.2, 150.1,146.6, 146.3, 139.4, 132.8, 132.3, 131.5, 130.0, 129.8, 118.1, 116.1,116.0, 115.2, 71.3, 71.3, 69.8, 62.7, 55.3, 36.3, 26.2, 26.1, 26.0,22.3, 21.3, 20.9, 18.7, −4.3, −5.4 ppm; HRMS (ESI-TOF) calcd forC₄₁H₅₇O₈Si₂ ⁺ [M+H]⁺ 733.3585, found 733.3605.

Epoxy Ketone 6 (Plus 4-Epi-6):

To a stirred solution of aldehyde 23 (800 mg, 1.07 mmol) in CHCl₃ (21mL) and H₂O (1.1 mL) at 25° C. were added urea.H₂O₂ (706 mg, 7.49 mmol,7.0 equiv) and (S)-(−)-α,α-diphenyl-2-pyrrolidine methanoltrimethylsilyl ether (24, 70.2 mg, 0.218 mmol, 0.2 equiv). Afterstirring at this temperature for 7 h, the reaction mixture was dilutedwith EtOAc (100 mL). The resulting mixture was washed with H₂O (2×50mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressureto give the crude epoxide 25, which was taken to the next step withoutfurther purification.

To a solution of the above crude epoxide 25 in THF (21 mL) at 25° C. wasadded DABCO (60.0 mg, 0.535 mmol, 0.5 equiv), 4-nitrophenol (74.8 mg,0.535 mmol, 0.5 equiv) and enone 7 (Edwards et al., 2003) (1.39 g, 10.7mmol, 10 equiv). After stirring at this temperature for 12 h, thereaction was diluted with EtOAc (100 mL). The resulting mixture waswashed with brine (2×50 mL) and concentrated under reduced pressure. Theresidue was purified by flash column chromatography (silica gel,EtOAc:hexanes 1:4-4:2) to give crude alcohol 26 containing a number ofunidentified side products.

The crude alcohol 26 obtained above was dissolved in CH₂Cl₂ (10 mL), andN-trimethylsilylimidazole (120 mg, 0.856 mmol, 0.8 equiv) was addedunder stirring at 0° C. The resulting reaction mixture was stirred atthis temperature for 30 min, and then concentrated to 2 mL under reducedpressure. The residue was purified by flash column chromatography(silica gel, EtOAc:CH₂Cl₂:hexanes 1:1:12) to give the title compound 6(plus C4-epi-6) (355 mg, 0.373 mmol, d.r. ca. 3:1, 36%, three steps) asa yellow foam. 6 (plus C4-epi-6): R_(f)=0.56 (silica gel, EtOAc:hexanes1:4); [α]²⁵ _(D)=+16.8 (c=1.0, CH₂Cl₂); FT-IR (neat): ν_(max)=2934,2898, 2859, 1696, 1614, 1560, 1514, 1471, 1445, 1371, 1250, 1158, 1055,1010, 938, 879, 829, 780, 662 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=7.36 (s,1H, major), 7.32-7.30 (m, 5H, major+minor), 6.87-6.84 (m, 4H,major+minor), 6.66 (s, 1H, major), 6.63 (s, 1H, minor), 6.48 (s, 1H,major), 6.45 (s, 1H, minor), 5.17-5.16 (m, 2H, major+minor), 5.11 (d,J=2.0 Hz, 1H, major), 5.08 (d, J=2.4 Hz, 1H, minor), 5.06 (s, 1H,major), 5.01 (s, 1H, minor), 4.86 (dd, J=12.4, 5.0 Hz, 2H, major+minor),4.71 (d, J=10.9 Hz, 2H, major+minor), 4.61 (d, J=10.8 Hz, 2H,major+minor), 4.11 (d, J=2.2 Hz, 1H, major), 3.89 (d, J=2.2 Hz, 1H,minor), 3.86 (s, 6H, major+minor), 3.79 (s, 6H, major+minor), 3.54 (t,J=2.5 Hz, 1H, minor), 3.50 (t, J=2.4 Hz, 1H, major), 3.42 (s, 6H,minor), 3.40 (s, 3H, major), 3.39 (s, 3H, major), 2.71 (ddd, J=13.5,4.9, 3.1 Hz, 2H, major+minor), 2.58 (s, 3H, major), 2.48 (s, 3H, minor),2.17-2.12 (m, 2H, major+minor), 1.18 (s, 9H, major), 1.15 (s, 9H,minor), 1.10 (s, 9H, minor), 1.10 (s, 9H, major), 0.95 (s, 18H,major+minor), 0.24 (s, 6H, major+minor), 0.15 (s, 9H, major), 0.14 (s,9H, minor), 0.14 (s, 6H, major+minor) ppm; ¹³C NMR (CDCl₃, 150 MHz)δ=194.4 (major+minor), 193.8 (minor), 193.8 (major), 159.3(major+minor), 152.1 (minor), 151.4 (major), 149.8 (major+minor), 146.1(major), 146.1 (minor), 144.2 (major), 144.0 (minor), 141.4 (major),140.5 (minor), 131.4 (major), 131.3 (minor), 130.4 (minor), 130.3(major), 130.3 (minor), 130.1 (major), 129.7 (major+minor), 129.3(major), 129.3 (minor), 119.7 (major), 119.6 (minor), 115.7 (minor),115.7 (major), 115.4 (major), 115.4 (minor), 114.3 (major), 114.2(minor), 113.8 (major+minor), 102.7 (minor), 102.1 (major), 71.3(major+minor), 71.2 (major+minor), 69.9 (minor), 69.8 (major), 67.8(minor), 67.1 (major), 62.6 (major+minor), 61.6 (major), 61.0 (minor),55.3 (major+minor), 54.6 (minor), 54.4 (minor), 54.3 (major), 54.1(major), 52.4 (major), 51.5 (minor), 36.5 (major), 36.5 (minor), 26.4(major+minor), 26.2 (minor), 26.2 (major), 26.0 (major+minor), 21.2(major), 21.2 (minor), 21.1 (minor), 21.1 (major), 21.0 (major), 20.9(minor), 18.7 (major+minor), 0.1 (major), 0.0 (minor), −4.3(major+minor), −5.4 (major+minor) ppm; HRMS (ESI-TOF) calcd forC₅₀H₇₅O₁₂Si₃ ⁺ [M+H]⁺ 951.4561, found 951.4581.

Acetal 27 (Stereochemistry Assigned by Coupling ConstantStudies^([11])):

To a stirred solution of epoxy ketone 6 (plus C4-epi-6) (20.1 mg, 0.021mmol) in CH₂Cl₂ (0.8 mL) at −78° C. was added SnCl₄ (0.01 M in CH₂Cl₂,21 μL, 0.021 mmol, 0.1 equiv) dropwise. After stirring at thistemperature for 2 h, the reaction was quenched with NaHCO₃ (sat. aq., 2mL). The resulting mixture was extracted with CH₂Cl₂ (3×2 mL), and thecombined organic phases were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography (silica gel, EtOAc:hexanes 1:13) to give the titlecompound (27, plus C4-epi-27) (7.3 mg, 0.0076 mmol, d.r. ca. 13:1, 37%)as yellow oil. R_(f)=0.58 (silica gel, EtOAc:hexanes 1:4); 27(+C4-epi-27): R_(f)=0.63 (silica gel, EtOAc:hexanes 1:4); [α]²⁵_(D)=+30.4 (c=0.45, CH₂Cl₂); FT-IR (neat): ν_(max)=2952, 2934, 2898,2859, 1697, 1613, 1562, 1514, 1471, 1445, 1370, 1252, 1159, 1088, 1059,1011, 954, 890, 830, 661 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz, major isomer)δ=7.40 (s, 1H), 7.30 (d, J=8.6 Hz, 2H), 6.86 (d, J=8.6 Hz, 2H), 5.69 (s,1H), 5.60 (d, J=2.2 Hz, 1H), 5.18 (t, J=2.6 Hz, 1H), 5.17-5.15 (m, 2H),4.86 (dd, J=12.3, 5.1 Hz, 1H), 4.71-4.68 (m, 2H), 4.64 (s, 1H), 4.60 (d,J=10.8 Hz, 1H), 3.88 (s, 3H), 3.79 (s, 3H), 3.53 (s, 3H), 3.50 (s, 3H),2.75-2.70 (m, 1H), 2.65 (s, 3H), 2.17-2.12 (m, 1H), 1.17 (s, 9H), 1.11(s, 9H), 0.95 (s, 9H), 0.24 (s, 3H), 0.16 (s, 9H), 0.14 (s, 3H) ppm; ¹³CNMR (CDCl₃, 150 MHz, major isomer) δ=194.5, 159.3, 151.7, 150.2, 147.3,146.0, 142.1, 131.6, 130.3, 129.7, 129.5, 120.3, 116.3, 116.0, 114.3,113.8, 108.6, 107.6, 104.5, 79.7, 72.9, 71.3, 71.1, 71.1, 69.8, 62.6,57.1, 56.9, 55.3, 36.6, 26.3, 26.2, 26.0, 22.3, 21.5, 20.9, 18.7, 0.1,−4.3, −5.4 ppm; HRMS (ESI-TOF) calcd for C₅₀H₇₅O₁₂Si₃ ⁺ [M+H]⁺ 951.4561,found 951.4576.

Acetal 5a and 5b:

To a stirred solution of epoxy ketone 6 (plus 4-epi-6) (140 mg, 0.147mmol) in CH₂Cl₂ (4.4 mL) at −78° C. was added BF₃.OEt₂ (0.1 M in CH₂Cl₂,440 μL, 0.044 mmol, 0.3 equiv) drop wise. After stirring at thistemperature for 6 h, the reaction was quenched sequentially with Et₃N(40 μL) and NaHCO₃ (sat. aq., 10 mL). The resulting mixture wasextracted with CH₂Cl₂ (3×10 mL), and the combined organic phases weredried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Theresidue was purified by flash column chromatography (silica gel,EtOAc:hexanes 1:20) to give the title compound (5a, 75.0 mg, 0.079 mmol,54%; 5b, 25.0 mg, 0.026 mmol, 18%) as a yellow foam.

5a (stereochemistry assigned by coupling constant studies [Padwa et al.,1991; Kraehenbuehl et al., 1995; Kraehenbuehl et al., 1998; Muthusamy etal., 2002]): R_(f)=0.68 (silica gel, EtOAc:hexanes 1:4); [α]²⁵_(D)=+168.8 (c=1.0, CH₂Cl₂); FT-IR (neat): ν_(max)=2934, 2897, 2859,1697, 1609, 1559, 1514, 1471, 1445, 1398, 1369, 1250, 1161, 1110, 1052,1032, 999, 892, 878, 827, 731, 661 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=7.31(s, 1H), 7.29 (d, J=9.1 Hz, 2H), 6.87 (d, J=8.6 Hz, 2H), 5.70 (d, J=3.1Hz, 1H), 5.47 (d, J=2.3 Hz, 1H), 5.23 (t, J=2.7 Hz, 1H), 5.06 (d, J=2.0Hz, 1H), 4.96 (dd, J=4.2, 3.0 Hz, 1H), 4.87-4.84 (m, 2H), 4.71-4.69 (m,2H), 4.60 (d, J=10.7 Hz, 1H), 3.89 (s, 3H), 3.79 (s, 3H), 3.62 (s, 3H),3.61 (s, 3H), 2.73 (ddd, J=13.5, 4.9, 3.1 Hz, 1H), 2.66 (s, 3H), 2.14(dt, J=13.4, 2.7 Hz, 1H), 1.14 (s, 9H), 1.06 (s, 9H), 0.96 (s, 9H), 0.24(s, 3H), 0.14 (s, 3H), −0.33 (s, 9H) ppm; ¹³C NMR (CDCl₃, 150 MHz)δ=194.4, 159.3, 149.3, 148.8, 146.7, 146.2, 141.6, 130.2, 130.1, 129.8,120.8, 116.8, 115.0, 113.9, 108.8, 106.6, 102.2, 79.9, 79.8, 73.2, 71.4,71.1, 70.1, 62.7, 56.2, 55.7, 55.3, 36.4, 26.5, 26.3, 26.0, 24.1, 21.7,20.5, 18.7, −0.4, −4.3, −5.4 ppm; HRMS (ESI-TOF) calcd forC₅₀H₇₄NaO₁₂Si₃ ⁺ [M+Na]⁺ 973.4370, found 973.4380.

5b (stereochemistry assigned by coupling constant studies [Padwa et al.,1991; Kraehenbuehl et al., 1995; Kraehenbuehl et al., 1998; Muthusamy etal., 2002]): R_(f)=0.65 (silica gel, EtOAc:hexanes 1:4); [α]²⁵_(D)=+165.8 (c=1.0, CH₂Cl₂); FT-IR (neat): ν_(max)=2935, 2860, 1698,1610, 1560, 1514, 1471, 1445, 1399, 1371, 1250, 1162, 1059, 1033, 1012,938, 885, 840, 828, 781, 662 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=7.34 (s,1H), 7.32 (d, J=8.5 Hz, 2H), 6.88 (d, J=8.5 Hz, 2H), 5.60 (s, 1H), 5.46(d, J=3.6 Hz, 1H), 5.20 (t, J=2.6 Hz, 1H), 5.16 (s, 1H), 5.04 (d, J=3.7Hz, 1H), 4.96 (s, 1H), 4.87 (dd, J=12.6, 4.8 Hz, 1H), 4.74 (d, J=10.7Hz, 1H), 4.64 (d, J=10.7 Hz, 1H), 4.17 (s, 1H), 3.90 (s, 3H), 3.79 (s,3H), 3.62 (s, 3H), 3.61 (s, 3H), 2.73 (ddd, J=13.5, 4.7, 3.2 Hz, 1H),2.59 (s, 3H), 2.15 (dt, J=13.4, 2.6 Hz, 1H), 1.14 (s, 9H), 1.06 (s, 9H),0.96 (s, 9H), 0.24 (s, 3H), 0.14 (s, 3H), −0.06 (s, 9H) ppm; ¹³C NMR(CDCl₃, 150 MHz) δ=194.5, 159.4, 150.2, 149.8, 149.2, 146.2, 139.9,130.5, 130.1, 129.9, 129.0, 119.2, 117.6, 115.3, 114.4, 113.9, 109.5,107.0, 101.4, 85.4, 77.4, 71.3, 71.2, 70.0, 62.7, 56.2, 55.3, 55.0,36.3, 26.4, 26.3, 26.0, 23.9, 21.7, 20.6, 18.7, 0.5, −4.3, −5.4 ppm;HRMS (ESI-TOF) calcd for C₅₀H₇₅O₁₂Si₂ ⁺ [M+H]⁺ 951.4561, found 951.4586.

Allylic Alcohol 28:

Acetal 5a (52.2 mg, 0.548 mmol) was dissolved in a solution of TFA (0.1M in THF:H₂O 5:1, 5.5 mL). After stirring at 25° C. for 5 h, thereaction was quenched with NaHCO₃ (sat. aq., 10 mL). The resultingmixture was extracted with CH₂Cl₂ (3×5 mL), and the combined organicphases were dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The residue was purified by flash column chromatography(silica gel, EtOAc:hexanes 1:10-4:4) to give the title compound (28,31.0 mg, 0.035 mmol, 65%) as a yellow foam and recovered startingmaterial (5a, 12.5 mg, 0.013 mmol, 24%). 28: R_(f)=0.61 (silica gel,EtOAc:hexanes 1:2); [α]²⁵ _(u)=+134.5 (c=1.0, CH₂Cl₂); FT-IR (neat):ν_(max)=3497, 2934, 2898, 2859, 1697, 1610, 1559, 1514, 1471, 1445,1398, 1372, 1249, 1162, 1087, 1053, 1033, 937, 827, 781, 661 cm⁻¹; ¹HNMR (CDCl₃, 600 MHz) δ=7.37 (s, 1H), 7.30 (d, J=8.6 Hz, 2H), 6.86 (d,J=8.6 Hz, 3H), 5.66 (d, J=3.0 Hz, 1H), 5.53 (d, J=2.4 Hz, 1H), 5.30 (d,J=2.4 Hz, 1H), 5.21-5.19 (m, 2H), 4.88 (dd, J=12.4, 5.0 Hz, 2H), 4.84(s, 1H), 4.72 (d, J=10.9 Hz, 1H), 4.65-4.64 (m, 2H), 4.61 (d, J=10.9 Hz,1H), 3.89 (s, 3H), 3.78 (s, 3H), 3.62 (s, 3H), 3.61 (s, 3H), 2.74-2.68(m, 1H), 2.68 (s, 3H), 2.14 (dt, J=13.6, 2.6 Hz, 1H), 1.15 (s, 9H), 1.09(s, 9H), 0.95 (s, 9H), 0.23 (s, 3H), 0.13 (s, 3H) ppm; ¹³C NMR (CDCl₃,150 MHz) δ=194.5, 159.3, 150.1, 149.1, 147.3, 146.1, 138.6, 130.8,130.2, 129.7, 129.3, 119.3, 118.3, 115.3, 114.5, 113.8, 108.7, 107.5,102.2, 79.5, 79.2, 75.2, 71.3, 71.2, 69.8, 62.7, 56.3, 55.8, 55.3, 36.4,26.2, 26.0, 24.7, 21.5, 20.9, 18.7, −4.3, −5.4 ppm; HRMS (ESI-TOF) calcdfor C₄₇H₆₇O₁₂Si₂ ⁺ [M+H]⁺ 879.4166, found 879.4177.

Epoxy Alcohol 29:

To a stirred solution of the allylic alcohol 28 (30.1 mg, 0.034 mmol) inacetone (1.0 mL) at 25° C. were sequentially added OsO₄ (0.08 M aq., 85μL, 0.068 mmol, 0.2 equiv) and NMO (0.48 M aq., 283 μL, 0.136 mmol, 4.0equiv). After stirring at this temperature for 12 h, the reaction wasquenched with Na₂SO₃ (10% aq., 10 mL). The resulting mixture was stirredfor another 30 min, then extracted with CH₂Cl₂ (3×5 mL), and thecombined organic phases were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography (silica gel, EtOAc:hexanes 1:2) to give the triolintermediate (26.3 mg, 0.029 mmol) as a yellow foam. To a stirredsolution of the above triol intermediate (26.3 mg, 0.029 mmol) in CH₂Cl₂(1.0 mL) at 25° C. were added Et₃N (14.6 mg, 0.144 mmol, 5.0 equiv),DMAP (2.0 mg, 0.0144 mmol, 0.5 equiv) and TsCl (16.4 mg, 0.144 mmol, 5.0equiv), sequentially. After stirring at this temperature for 5 h, thereaction was quenched with NH₄Cl (sat. aq., 5 mL). The resulting mixturewas extracted with CH₂Cl₂ (3×5 mL), and the combined organic phases weredried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Theresidue was purified by flash column chromatography (silica gel,EtOAc:hexanes 1:2) to give the corresponding primary tosylateintermediate (31.0 mg, 0.029 mmol) as a yellow foam.

To a stirred solution of the above tosylate intermediate (31.0 mg, 0.029mmol) in MeOH (1.0 mL) at 25° C. was added K₂CO₃ (8.0 mg, 0.058 mmol,2.0 equiv). The resulting reaction mixture was stirred at thistemperature for 1 h and was then directly subjected to flash columnchromatography (silica gel, EtOAc:hexanes 4:1) to give the titlecompound (29, 25.0 mg, 0.028 mmol, 82%, three steps) as a yellow foam.29: R_(f)=0.52 (silica gel, EtOAc:hexanes 1:2); [α]²⁵ _(D)=+106.5(c=1.0, CH₂Cl₂); FT-IR (neat): ν_(max)=3487, 2934, 2859, 1696, 1610,1560, 1514, 1471, 1445, 1399, 1371, 1249, 1162, 1079, 1055, 1003, 978,828, 662 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=7.39 (s, 1H), 7.29 (d, J=8.6Hz, 2H), 6.85 (d, J=8.6 Hz, 2H), 5.66 (d, J=2.8 Hz, 1H), 5.40 (dd,J=4.8, 3.3 Hz, 1H), 5.19 (t, J=2.5 Hz, 1H), 4.86 (dd, J=12.3, 5.1 Hz,1H), 4.73 (s, 1H), 4.71 (d, J=10.9 Hz, 1H), 4.60 (d, J=10.9 Hz, 1H),4.31 (dd, J=9.1, 5.1 Hz, 1H), 3.89 (s, 3H), 3.78 (s, 3H), 3.63 (s, 3H),3.55 (s, 3H), 3.29 (d, J=5.5 Hz, 1H), 3.07 (d, J=5.4 Hz, 1H), 2.73-2.71(m, 4H), 2.14 (dt, J=13.6, 2.5 Hz, 1H), 1.15 (s, 9H), 1.09 (s, 9H), 0.95(s, 9H), 0.23 (s, 3H), 0.13 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz)δ=194.5, 159.2, 150.0, 149.1, 146.1, 138.9, 130.7, 130.2, 129.6, 129.4,118.7, 118.1, 115.3, 114.6, 113.8, 108.0, 100.1, 79.3, 78.0, 76.2, 71.3,71.1, 69.7, 67.9, 62.7, 56.7, 56.1, 55.3, 47.6, 36.5, 26.3, 26.3, 26.0,24.0, 21.5, 20.8, 18.7, −4.3, −5.4 ppm; HRMS (ESI-TOF) calcd forC₄₇H₆₇O₁₃Si₂ ⁺ [M+H]⁺ 895.4136, found 895.4115.

Keto Epoxide 30:

To a stirred solution of the epoxy alcohol 29 (30.0 mg, 0.034 mmol) inCH₂Cl₂ at 25° C. were added NMO.H₂O (13.6 mg, 0.101 mmol, 3.0 equiv) andTPAP (2.4 mg, 0.007 mmol, 0.2 equiv). The resulting reaction mixture wasstirred at this temperature for 1 h, and then directly subjected toflash column chromatography (silica gel, EtOAc:hexanes 1:4) to give thetitle compound (30, 27.8 mg, 0.031 mmol, 93%) as a yellow foam. 30:R_(f)=0.51 (silica gel, EtOAc:hexanes 1:4); [α]²⁵ _(D)=+192.9 (c=1.0,CH₂Cl₂); FT-IR (neat): ν_(max)=2934, 2859, 1788, 1702, 1611, 1514, 1471,1445, 1373, 1250, 1162, 1082, 1045, 1010, 827, 662 cm⁻¹; ¹H NMR (CDCl₃,600 MHz) δ=7.33 (s, 1H), 7.29 (d, J=8.5 Hz, 2H), 6.86 (d, J=8.5 Hz, 2H),5.62 (d, J=3.8 Hz, 1H), 5.45 (d, J=3.8 Hz, 1H), 5.19 (br s, 1H),4.88-4.85 (m, 2H), 4.70 (d, J=10.9 Hz, 1H), 4.60 (d, J=10.9 Hz, 1H),3.87 (s, 3H), 3.79 (s, 3H), 3.69 (s, 3H), 3.58 (s, 3H), 3.39 (d, J=6.4Hz, 1H), 3.10 (d, J=6.4 Hz, 1H), 2.74-2.70 (m, 1H), 2.59 (s, 3H),2.15-2.11 (m, 1H), 1.17 (s, 9H), 1.09 (s, 9H), 0.95 (s, 9H), 0.23 (s,3H), 0.13 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=202.1, 194.5, 159.2,150.3, 149.1, 146.1, 138.8, 131.0, 130.2, 129.7, 129.6, 118.0, 117.1,115.3, 114.6, 113.8, 107.5, 99.9, 82.0, 78.0, 71.3, 71.1, 69.7, 62.7,61.8, 57.1, 56.2, 55.3, 50.1, 36.4, 26.3, 26.2, 26.0, 23.8, 21.5, 20.8,18.7, −4.3, −5.4 ppm; HRMS (ESI-TOF) calcd for C₄₇H₆₅O₁₃Si₂ ⁺ [M+H]⁺893.3958, found 893.3934. All spectroscopic data were consistent withthose reported in the literature. (S̆venda et al., 2011)

Hemiacetal 31:

To a stirred solution of keto epoxide 30 (20.1 mg, 0.0225 mmol) in CH₃CN(1.0 mL) at 25° C. was added Et₃N.3HF (10.0 mg, 0.061 mmol, 3.0 equiv).After stirring at this temperature for 15 min, the reaction was quenchedwith NaHCO₃ (5% aq., 5 mL) and diluted with EtOAc (10 mL). The resultingmixture was washed sequentially with H₂O (5 mL) and brine (5 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure. Theresidue was purified by flash column chromatography (silica gel,EtOAc:hexanes 2:1) to give the title compound (31, 15.0 mg, 0.0199 mmol,88%) as an orange foam. 31: R_(f)=0.42 (silica gel, EtOAc:hexanes 2:1);[α]²⁵ _(D)=+200.3 (c=1.0, CH₂Cl₂); FT-IR (neat): ν_(max)=3412, 2952,2930, 2855, 1620, 1570, 1514, 1390, 1124, 1067, 1033, 983, 945, 870,836, 778 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=14.78 (s, 1H), 7.41 (s, 1H),7.27 (d, J=8.6 Hz, 2H), 6.86 (d, J=8.6 Hz, 2H), 5.25 (d, J=4.0 Hz, 1H),5.16 (t, J=2.5 Hz, 1H), 4.94 (dd, J=12.4, 5.1 Hz, 1H), 4.88 (d, J=4.0Hz, 1H), 4.69 (s, 1H), 4.69 (d, J=11.0 Hz, 1H), 4.58 (d, J=11.0 Hz, 1H),4.48 (br s, 1H), 3.81 (s, 3H), 3.79 (s, 3H), 3.61 (s, 3H), 3.46 (s, 3H),3.09 (d, J=5.3 Hz, 1H), 2.95 (d, J=5.3 Hz, 1H), 2.71 (ddd, J=13.4, 5.0,3.4 Hz, 1H), 2.57 (s, 3H), 2.18 (td, J=13.4, 2.3 Hz, 1H), 0.96 (s, 9H),0.23 (s, 3H), 0.16 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=203.3, 162.6,159.3, 151.4, 144.1, 141.8, 135.1, 130.1, 129.5, 127.6, 116.4, 114.9,114.6, 113.9, 108.6, 103.8, 100.1, 98.5, 73.2, 70.9, 69.7, 69.3, 69.2,68.9, 62.7, 57.0, 56.7, 55.3, 50.4, 36.2, 25.9, 20.4, 18.6, −4.4, −5.3ppm; HRMS (ESI-TOF) calcd for C₃₉H₄₉O₁₃Si⁺ [M+H]⁺ 753.2937, found753.2952. All spectroscopic data were consistent with those reported inthe literature. (S̆venda et al., 2011)

Hydroxy Hemiacetal 32:

To a stirred solution of hemiacetal 31 (15.2 mg, 0.0202 mmol) in CH₂Cl₂(1.0 mL) and H₂O (0.1 mL) at 25° C. in a reaction flask shielded fromlight using aluminum foil was added DDQ (6.9 mg, 0.0303 mmol, 1.5equiv). After stirring at this temperature for 3 h, the reaction wasquenched with brine (5 mL). The resulting mixture was extracted withCH₂Cl₂ (3×5 mL), dried over anhydrous Na₂SO₄, and concentrated underreduced pressure. The residue was purified by flash columnchromatography (silica gel, EtOAc:hexanes 3:1) to give the titlecompound (31, 11.9 mg, 0.0187 mmol, 93%) as an orange foam. 32:R_(f)=0.70 (silica gel, EtOAc); [α]²⁵ _(D)=+131.5 (c=0.2, CH₂Cl₂); FT-IR(neat): ν_(max)=2930, 2854, 1620, 1570, 1514, 1444, 1390, 1250, 1157,1124, 1067, 983, 871, 837, 779 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=14.63 (s,1H), 7.43 (s, 1H), 5.42 (t, J=3.0 Hz, 1H), 5.25 (d, J=4.0 Hz, 1H), 4.88(dd, J=11.7, 4.9 Hz, 1H), 4.83 (d, J=4.0 Hz, 1H), 4.70 (s, 1H), 4.42 (brs, 1H), 3.91 (s, 3H), 3.62 (s, 3H), 3.47 (s, 3H), 3.13 (d, J=5.3 Hz,1H), 3.03 (d, J=5.3 Hz, 1H), 2.60 (s, 3H), 2.53 (br s, 1H), 2.49 (dt,J=13.5, 4.5 Hz, 1H), 2.34 (dt, J=13.6, 3.4 Hz, 1H), 0.94 (s, 9H), 0.21(s, 3H), 0.16 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=202.8, 162.4,151.5, 143.8, 142.0, 135.2, 129.6, 116.1, 114.8, 114.6, 107.9, 103.9,100.1, 98.5, 73.2, 69.6, 69.4, 69.3, 62.6, 57.0, 56.6, 50.5, 38.6, 25.8,20.4, 18.5, 4.5, −5.3 ppm; HRMS (ESI-TOF) calcd for C₃₁H₄₁O₁₂Si⁺[M+H]⁺633.2362, found 633.2344.

DC-45-A2 (1):

To a stirred solution of ketone 31 (10.2 mg, 0.0161 mmol) in CH₃CN (1.0mL) at 25° C. in a reaction flask shielded from light using aluminumfoil was added Et₃N.3HF (49.0 mg, 0.30 mmol, 20 equiv). After stirringat this temperature for 12 h, the reaction was quenched with NaHCO₃ (5%aq., 5 mL) and diluted with EtOAc (10 mL). The resulting mixture waswashed sequentially with water (5 mL) and brine (5 mL), dried overanhydrous Na₂SO₄, and concentrated under reduced pressure. The residuewas purified by preparatory HPLC (Atlantis Prep T3 OBD column, 5 μm,19×150 mm, UV detection at 271 nm, isocratic elution with 20% MeCN inH₂O, flow rate: 10 mL/min, 32→34 min) to give DC-45-A2 (1, 7.2 mg,0.0138 mmol, 86%) as an orange solid. 1: R_(f)=0.21-0.62 (tailing,silica gel, EtOAc); [α]²⁵ _(D)=+182 (c=0.3, CH₂Cl₂); FT-IR (neat):ν_(max)=3364, 2961, 2926, 2853, 1621, 1571, 1446, 1387, 1099, 1067,1014, 940, 801 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=14.00 (s, 1H), 7.46 (s,1H), 5.45 (s, 1H), 5.26 (d, J=3.9 Hz, 1H), 4.92 (dd, J=12.6, 5.2 Hz,1H), 4.84 (d, J=3.8 Hz, 1H), 4.71 (s, 1H), 3.92 (s, 3H), 3.62 (s, 3H),3.47 (s, 3H), 3.15 (d, J=5.4 Hz, 1H), 3.03 (d, J=5.3 Hz, 1H), 2.74-2.70(m, 1H), 2.62 (s, 3H), 2.21-2.16 (m, 1H) ppm; ¹³C NMR (CDCl₃, 150 MHz)δ=203.2, 162.3, 151.7, 144.4, 142.7, 135.7, 129.4, 116.4, 115.0, 114.7,107.1, 103.9, 100.1, 98.6, 73.2, 69.6, 69.3, 67.7, 62.8, 61.9, 57.0,56.7, 50.5, 36.9, 20.5 ppm; HRMS (ESI-TOF) calcd for C₂₅H₂₇O₁₂ ⁺ [M+H]⁺519.1497, found 519.1482. All spectroscopic data were consistent withthose reported in the literature. (S̆venda et al., 2011)

KCN-Trox5:

[α]²⁵ _(D)=+190.9 (c=0.23, CH₂Cl₂); FT-IR (neat): ν_(max)=2926, 2852,1622, 1570, 1446, 1390, 1225, 1193, 1113, 1078, 1043, 1006, 974, 801cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=15.04 (s, 1H), 7.42 (s, 1H), 5.22 (d,J=4.1 Hz, 1H), 4.81 (d, J=4.1 Hz, 1H), 4.75 (s, 1H), 3.79 (s, 3H), 3.74(s, 3H), 3.63 (s, 3H), 3.45 (s, 3H), 3.05 (dd, J=5.0, 7.2 Hz, 2H), 2.90(d, J=5.7 Hz, 1H), 2.88 (d, J=5.7 Hz, 1H), 2.59 (s, 3H), 1.95 (dd,J=6.4, 6.4 Hz, 2H), 1.30 (s, 3H), 1.29 (s, 3H) ppm; ¹³C NMR (CDCl₃, 150MHz) δ=209.5, 163.5, 151.4, 142.2, 141.6, 135.1, 129.9, 115.5, 113.4,113.1, 109.5, 104.5, 102.0, 99.9, 71.4, 69.2, 60.8, 57.1, 56.7, 52.8,41.6, 35.6, 25.1, 25.0, 20.4, 19.6 ppm.

KCN-Trox6:

[α]²⁵ _(D)=+201.7 (c=0.18, CH₂Cl₂); FT-IR (neat): ν_(max)=2926, 1767,1622, 1569, 1446, 1391, 1211, 1089, 1044, 990, 868, 803 cm⁻¹; ¹H NMR(CDCl₃, 600 MHz) δ=14.88 (s, 1H), 7.43 (s, 1H), 5.60 (d, J=4.1 Hz, 1H),5.26 (d, J=4.2 Hz, 1H), 4.75 (s, 1H), 3.78 (s, 3H), 3.63 (s, 3H), 3.47(s, 3H), 3.05-3.01 (m, 3H), 2.96 (d, J=5.8 Hz, 1H), 2.58 (s, 3H), 2.26(s, 3H), 1.96-1.89 (m, 1H), 1.28 (s, 3H), 1.26 (s, 3H) ppm; ¹³C NMR(CDCl₃, 150 MHz) δ=209.3, 169.0, 163.5, 150.9, 142.1, 141.3, 135.1,130.0, 115.8, 113.1, 112.6, 109.6, 103.9, 100.9, 99.7, 71.1, 69.3, 69.1,60.8, 56.8, 56.2, 48.0, 41.5, 35.6, 25.1, 25.0, 21.8, 20.3, 19.6 ppm.

KCN-Trox8:

R_(f)=0.77 (silica gel, EtOAc:hexanes 1:1); [α]²⁵ _(D)=+268.8 (c=0.08,CH₂Cl₂); FT-IR (neat): ν_(max)=2920, 1620, 1570, 1445, 1389, 1236, 1180,1094, 1074, 1014, 918 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=14.80 (s, 1H),7.43 (s, 1H), 5.26 (d, J=4.1 Hz, 1H), 5.08 (s, 1H), 4.84 (d, J=4.2 Hz,1H), 4.29 (dd, J=4.8, 11.4 Hz, 1H), 4.12 (dd, J=4.8, 11.4 Hz, 1H), 3.89(dt, J=3.0, 12.6 Hz, 1H), 3.80-3.76 (m, 4H), 3.74 (s, 3H), 3.08-2.99 (m,1H), 2.96 (d, J=6.0 Hz, 1H), 2.87 (d, J=5.4 Hz, 1H), 2.73 (t, J=6.6 Hz,2H), 2.61 (s, 3H), 2.26-2.17 (m, J=1H), 2.11-2.07 (m, J=2H), 1.37 (d,J=13.7 Hz, 1H) ppm; ¹³C NMR (CDCl₃, 150 MHz) δ=204.4, 163.01, 151.5,142.4, 142.0, 135.2, 130.2, 115.6, 113.2, 113.2, 111.0, 103.1, 102.0,96.3, 71.5, 69.2, 69.0, 67.5, 67.4, 60.9, 52.8, 38.8, 25.6, 23.6, 22.1,20.7 ppm.

KCN-Trox9:

[α]²⁵ _(D)=+181.8 (c=0.11, CH₂Cl₂); FT-IR (neat): ν_(max)=2923, 1767,1621, 1571, 1445, 1389, 1347, 1234, 1212, 1180, 1097, 1010, 984, 923,876 cm⁻¹; ¹H NMR (CDCl₃, 600 MHz) δ=14.80 (s, 1H), 7.44 (s, 1H), 5.62(d, J=4.1 Hz, 1H), 5.30 (d, J=4.1 Hz, 1H), 5.08 (s, 1H), 4.30 (dd,J=2.4, 11.8 Hz, 1H), 4.15 (dd, J=4.9, 11.5 Hz, 1H), 3.90 (dt, J=2.5,12.2 Hz, 1H), 3.79 (dt, J=2.5, 12.1 Hz, 1H), 3.76 (s, 3H), 3.74 (s, 3H),3.03-2.98 (m, 4H), 2.70 (t, J=6.4 Hz, 2H), 2.60 (s, 3H), 2.26 (s, 3H),2.25-2.19 (m, J=1H), 2.09-2.05 (m, J=2H), 1.39 (d, J=13.6 Hz, 1H) ppm;¹³C NMR (CDCl₃, 150 MHz) δ=204.3, 168.9, 163.0, 151.0, 142.3, 141.7,135.3, 130.3, 116.0, 113.0, 112.7, 111.1, 102.6, 110.8, 96.3, 71.2,69.5, 69.0, 67.5, 67.4, 60.9, 48.0, 38.8, 25.5, 23.6, 22.1, 21.8, 20.6ppm.

Example 4—¹H and ¹³C NMR Structural Comparison to S̆venda, et al

Comparison of ¹H and ¹³C NMR Spectroscopic Data of Keto Epoxide 30(S̆venda, et al. and Herein)

TABLE 1a Comparison of ¹H NMR spectroscopic data Data from {hacek over(S)}venda, et al. Data from this work Δδ (CDCl₃, 500 MHz) (CDCl₃, 600MHz) (ppm) 7.33 (s, 1 H) 7.33 (s, 1 H) 0.00 7.29 (d, J = 8.5 Hz, 2 H)7.29 (d, J = 8.5 Hz, 2 H) 0.00 6.86 (d, J = 9.0 Hz, 2 H) 6.86 (d, J =8.5 Hz, 2 H) 0.00 5.62 (d, J = 4.0 Hz, 1 H) 5.62 (d, J = 3.8 Hz, 1 H)0.00 5.44 (d, J = 4.0 Hz, 1 H) 5.45 (d, J = 3.8 Hz, 1 H) −0.01  5.19(dd, J = 3.0, 2.5 Hz, 1 H) 5.19 (brs, 1 H) 0.00 4.86 (dd, J = 12.0, 5.0Hz, 1 H) 4.88-4.85 (m, 2H) — 4.85 (s, 1 H) 4.70 (d, J = 10.5 Hz, 1 H)4.70 (d, J = 10.9 Hz, 1 H) 0.00 4.60 (d, J = 11.0 Hz, 1 H) 4.60 (d, J =10.9 Hz, 1 H) 0.00 3.87 (s, 3 H) 3.87 (s, 3H) 0.00 3.79 (s, 3 H) 3.79(s, 3 H) 0.00 3.69 (s, 3 H) 3.69 (s, 3 H) 0.00 3.58 (s, 3 H) 3.58 (s, 3H) 0.00 3.39 (d, J = 7.0 Hz, 1 H) 3.39 (d, J = 6.4 Hz, 1 H) 0.00 3.10(d, J = 6.0 Hz, 1 H) 3.10 (d, J = 6.4 Hz, 1 H) 0.00 2.74-2.70 (m, 1 H)2.74-2.70 (m, 1 H) 0.00 2.59 (s, 3 H) 2.59 (s, 3 H) 0.00 2.16-2.10 (m, 1H) 2.15-2.11 (m, 1 H) — 1.17 (s, 9 H) 1.17 (s, 9 H) 0.00 1.09 (s, 9 H)1.09 (s, 9 H) 0.00 0.95 (s, 9 H) 0.95 (s, 9 H) 0.00 0.23 (s, 3 H) 0.23(s, 3 H) 0.00 0.13 (s, 3 H) 0.13 (s, 3 H) 0.00

TABLE 1b Comparison of ¹³C NMR spectroscopic data Data from {hacek over(S)}venda, et al. Data from this work Δδ (CDCl₃, 125 MHz) (CDCl₃, 150MHz) (ppm) 202.1 202.1 0.0 194.5 194.5 0.0 159.3 159.2 0.1 150.3 150.30.0 149.1 149.1 0.0 146.1 146.1 0.0 138.8 138.8 0.0 131.0 131.0 0.0130.3 130.2 0.1 129.7 129.7 0.0 129.6 129.6 0.0 118.0 118.0 0.0 117.1117.1 0.0 115.3 115.3 0.0 114.6 114.6 0.0 113.8 113.8 0.0 107.5 107.50.0 99.9 99.9 0.0 82.0 82.0 0.0 78.0 78.0 0.0 71.3 71.3 0.0 71.1 71.10.0 69.7 69.7 0.0 62.8 62.7 0.1 61.8 61.8 0.0 57.1 57.1 0.0 56.2 56.20.0 55.3 55.3 0.0 50.1 50.1 0.0 36.4 36.4 0.0 26.3 26.2 0.1 26.2 26.20.0 26.0 26.0 0.0 23.8 23.8 0.0 21.5 21.5 0.0 20.9 20.8 0.1 18.7 18.70.0 −4.3 −4.3 0.0 −5.4 −5.4 0.0Comparison of ¹H and ¹³C NMR Spectroscopic Data of Hemiacetal 31(S̆venda, et al. and Herein)

TABLE 2a Comparison of ¹H NMR spectroscopic data Data from {hacek over(S)}venda, et al. Data from this work Δδ (CDCl₃, 500 MHz) (CDCl₃, 600MHz) (ppm) 14.81 (s, 1 H) 14.78 (s, 1 H) 0.03 7.47 (s, 1 H) 7.41 (s, 1H) 0.06 7.28 (d, J = 8.5 Hz, 2 H) 7.27 (d, J = 8.6 Hz, 2 H) 0.01 6.87(d, J = 9.0 Hz, 2 H) 6.86 (d, J = 8.6 Hz, 2 H) 0.01 5.26 (d, J = 4.0 Hz,1 H) 5.25 (d, J = 4.0 Hz, 1 H) 0.01 5.18 (dd, J = 3.0, 2.0 Hz, 1 H) 5.16(t, J = 2.5 Hz, 1 H) 0.02 4.95 (dd, J = 12.5, 5.0 Hz, 1 H) 4.94 (dd, J =0.01 12.4, 5.1 Hz, 1 H) 4.84 (d, J = 4.0 Hz, 1 H) 4.88 (d, J = 4.0 Hz, 1H) −0.04  4.69 (d, J = 10.5 Hz, 1 H) 4.69 (d, J = 11.0 Hz, 1 H) 0.004.70 (s, 1 H) 4.69 (s, 1 H) 0.01 4.58 (d, J = 11.5 Hz, 1 H) 4.58 (d, J =11.0 Hz, 1 H) 0.00 4.31 (s, 1 H) 4.48 (s, 1 H) −0.17  3.83 (s, 3 H) 3.81(s, 3 H) 0.02 3.80 (s, 3 H) 3.79 (s, 3 H) 0.01 3.62 (s, 3 H) 3.61 (s, 3H) 0.01 3.46 (s, 3 H) 3.46 (s, 3 H) 0.00 2.98 (d, J = 5.5 Hz, 1 H) 2.95(d, J = 5.3 Hz, 1 H) 0.03 2.74-2.69 (m, 1 H) 2.71 (ddd, J = 13.4, — 5.0,3.4 Hz, 1 H) 2.60 (s, 3 H) 2.57 (s, 3 H) 0.03 2.20-2.15 (m, 1 H) 2.18(dt, J = — 13.4, 2.3 Hz, 1 H) 0.96 (s, 9 H) 0.96 (s, 9 H) 0.00 0.23 (s,3 H) 0.23 (s, 3 H) 0.00 0.16 (s, 3 H) 0.16 (s, 3 H) 0.00

TABLE 2b Comparison of ¹³C NMR spectroscopic data Data from {hacek over(S)}venda, et al. Data from this work Δδ (CDCl₃, 125 MHz) (CDCl₃, 150MHz) (ppm) 203.2 203.3 −0.1 162.6 162.6 0.0 159.3 159.3 0.0 151.3 151.4−0.1 144.0 144.1 −0.1 141.8 141.8 0.0 135.0 135.1 −0.1 130.1 130.0 0.1129.5 129.5 0.0 127.5 127.6 −0.1 116.3 116.4 −0.1 114.8 114.9 −0.1 114.7114.6 0.1 113.8 113.9 −0.1 108.5 108.6 −0.1 103.8 103.8 0.0 100.1 100.10.0 98.5 98.5 0.0 73.2 73.2 0.0 70.9 70.9 0.0 69.7 69.7 0.0 69.2 69.3−0.1 69.2 69.2 0.0 68.9 68.9 0.0 62.7 62.7 0.0 56.9 57.0 −0.1 56.6 56.7−0.1 55.2 55.3 −0.1 50.3 50.4 −0.1 36.1 36.2 −0.1 25.9 25.9 0.0 20.420.4 0.0 18.6 18.6 0.0 −4.4 −4.4 0.0 −5.4 −5.4 0.0Comparison of ¹H and ¹³C NMR Spectroscopic Data of DC-45-A2 (S̆venda, etal. and Herein)

TABLE 3a Comparison of ¹H NMR spectroscopic data Data from {hacek over(S)}venda, et al. Data from this work Δδ (CDCl₃, 500 MHz) (CDCl₃, 600MHz) (ppm) 13.99 (s, 1 H) 14.00 (s, 1 H) −0.01 7.43 (s, 1 H) 7.46 (s, 1H) −0.03 5.45 (s, 1 H) 5.45 (s, 1 H) 0.00 5.25 (d, J = 3.6 Hz, 1 H) 5.26(d, J = 3.9 Hz, 1 H) −0.01 4.91 (dd, J = 4.92 (dd, J = −0.01 12.6, 4.8Hz, 1 H) 12.6, 5.2 Hz, 1 H) 4.85 (d, J = 3.6 Hz, 1 H) 4.84 (d, J = 3.8Hz, 1 H) 0.01 4.71 (s, 1 H) 4.71 (s, 1 H) 0.00 4.58 (br s, 1 H) — — 3.92(s, 3 H) 3.92 (s, 3 H) 0.00 3.62 (s, 3 H) 3.62 (s, 3 H) 0.00 3.47 (s, 3H) 3.47 (s, 3 H) 0.00 3.13 (d, J = 4.8 Hz, 1 H) 3.15 (d, J = 5.4 Hz, 1H) −0.02 3.02 (d, J = 5.4 Hz, 1 H) 3.03 (d, J = 5.4 Hz, 1 H) −0.012.74-2.70 (m, 1 H) 2.74-2.70 (m, 1 H) — 2.61 (s, 3 H) 2.62 (s, 3 H)−0.01 2.33 (br s, 1 H) — — 2.21-2.17 (m, 1 H) 2.21-2.16 (m, 1 H) —

TABLE 3b Comparison of ¹³C NMR spectroscopic data Data from {hacek over(S)}venda, et al. Data from this work Δδ (CDCl₃, 125 MHz) (CDCl₃, 150MHz) (ppm) 203.3 203.2 0.1 162.2 162.3 −0.1 151.6 151.7 −0.1 144.4 144.40.0 142.6 142.7 −0.1 135.6 135.7 −0.1 129.3 129.4 −0.1 116.3 116.4 −0.1115.0 115.0 0.0 114.5 114.7 −0.2 107.2 107.1 0.1 103.9 103.9 0.0 100.1100.1 0.1 98.7 98.6 0.0 73.3 73.2 0.1 69.5 69.6 −0.1 69.2 69.3 −0.1 67.767.7 0.0 62.9 62.8 0.1 61.8 61.9 −0.1 57.0 57.0 0.0 56.6 56.7 −0.1 50.350.5 −0.2 37.0 36.9 0.1 20.5 20.5 0.0

Example 5—Biological Activity

A. Cytotoxity Assay

Cells were cultured in a T75 flask to ˜50-80% confluency and harvestedwith trypsin into a single cell suspension. Five hundred (500) cells perwell were seeded in tissue culture plates in 50 μL/well culture mediaand incubated at 37° C. for 18-24 hours. Compounds were diluted as 400×final desired concentrations in DMSO. Serial dilutions in DMSO were thendiluted in culture media for a final DMSO concentration of 0.25% and 50μL/well of the final dilution was added to the cells (Vf=100 μL). Uponplating and treatment, cells were returned to the incubator for anadditional 72 hours. CellTiter-Glo reagent was prepared permanufacturer's instructions and added at 100 μL/well to the cultures.CellTiter-Glo allows for relative enumeration of metabolically activecells by quantifying intracellular ATP concentrations. After 5 minutesof incubation with CellTiter-Glo at ambient room temperature, 125μL/well of the Cell Titer Glo/cell lysate solution was transferred intoblack assay plates, which were then read in a luminometer within 30minutes. Luminescence readings obtained from cultures that did notreceive any treatment (cell culture media only) were set as 100% controland all other luminescence values were normalized to these controls(e.g., Normalized RLU, relative luminescence unit).

B. Cell Lines Used in the Assay

MES SA and MES SA/Dx cells are uterine sarcoma. MES SA Dx cell line wasgenerated from MES SA to achieve upregulation of MDR1. MES-SA/Dx cellsexhibit marked cross-resistance to a number of chemotherapeutic agents(including daunorubicin, dactinomycin, vincristine, taxol, colchicine)and moderate cross-resistance to mitomycin C and melphalan. 293T cellsare a human embryonic kidney cell line.

C. Activity Results

The results of the assay are shown in FIGS. 3A-3C and 4A-4C and Table 4below. In these assays, Trox8 showed 530 pM activity in the MES SAassay, 380 pM in the MES SA DX assay, and 550 pM activity in the 293Tassay. Additionally, Trox5, Trox7, and Trox9 also showed good nanomolarcytotoxicity.

TABLE 4 Biological Activity of Trox4-Trox9 MES SA MES SA DX 293TCompound ID IC₅₀ nM IC₅₀ nM IC₅₀ nM

n/a

n/a

n/a

>1000 >1000 >1000

562.4 213 786.2

>1000 >1000 >1000

3.72 5.72 2.46

0.53 0.38 0.55

6.109 17.42 6.89

18.08 >1000 14.89

0.74 203.5 0.702

11.06 >1000 8.016

157.4 >1000 95.44

2.02 >1000 2.815

All of the compositions and/or methods disclosed and claimed herein canbe made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions and/or methods and in the steps or in the sequence ofsteps of the method described herein without departing from the concept,spirit and scope of the invention. More specifically, it will beapparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the spirit, scope and concept of theinvention as defined by the appended claims.

VIII. REFERENCES

The following references, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference:

-   Anderson, N. G., Practical Process Research & Development—A Guide    For Organic Chemists, 2^(nd) ed., Academic Press, New York, 2012.-   March's Advanced Organic Chemistry: Reactions, Mechanisms, and    Structure, 2007.-   Greene's Protective Groups in Organic Chemistry, Wuts and Greene,    Ed., 1973-   Remington's Pharmaceutical Sciences, 15^(th) Ed., 1035-1038 and    1570-1580, 1990.-   Remington's Pharmaceutical Sciences, 15^(th) Ed., 3:624-652, 1990.-   U.S. Pat. No. 4,459,291-   U.S. Pat. No. 5,739,169-   U.S. Pat. No. 5,801,005-   U.S. Pat. No. 5,824,311-   U.S. Pat. No. 5,830,880-   U.S. Pat. No. 5,846,945-   WO 2003/035065-   WO 2011/119549-   Banwell et al., J. Org. Chem. 1994, 59, 6338-6343.-   J. Cassidy et al., Cancer Chemother. Pharmacol. 1993, 31, 395-400.-   Colman et al., Tetrahedron: Asymmetry 1999, 10, 4175-4182.-   Davidson et al., J. Immunotherapy, 1998, 21(5):389.-   de Sousa et al., Tetrahedron 2002, 58, 4643-4654.-   Evans et al., J. Am. Chem. Soc. 1991, 113, 7613-7630.-   Fitzner et al., Anal. Bioanal. Chem. 2008, 390, 1139-1147.-   Gaoni, J. Chem. Soc. (C) 1968, 2925-2934.-   Hauser et al. Org. Chem. 1978, 43, 178-180.-   Hoye et al., Nat. Protoc. 2007, 10, 2451-2457.-   Kato et al., Tetrahedron 62, 7307-7318, 2006.-   Kraehenbuehl et al., Tetrahedron Lett. 1995, 36, 8595-8598;-   Kraehenbuehl et al., Helv. Chim. Acta 1998, 81, 1439-1479;-   Kraus et al., Tetrahedron Lett. 1978, 19, 2263-2266.-   Pfoh et al., Nucleic Acids Res. 2008, 36, 3508-3514.-   Magauer et al., Nat. Chem. 2013, 5, 886-893.-   Maiese et al., J. Antibiot. 1990, 43, 253-258.-   Maras et al., J. Org. Chem. 1998, 63, 2039-2041.-   Mango et al., J. Am. Chem. Soc. 2005, 127, 6964-6965.-   Maskey et al., J. Antibiot. 2004, 57, 771-779.-   Maskey et al., Angew. Chem. Int. Ed. 2004, 43, 1281-1283; Angew.    Chem. 2004, 116, 1301-1303.-   Muthusamy et al., J. Org. Chem. 2002, 67, 8019-8033.-   Naruse et al., Tetrahedron 1988a, 44, 4747-4756.-   Naruse et al., Tetrahedron Lett. 1988b, 29, 1417-1420.-   Nicolaou et al., J. Am. Chem. Soc. 2009, 131, 14812-14826.-   O'Brien et al., J. Chem. Soc., Perkin Trans. 1 1998, 2435-2441.-   Padwa et al., J. Org. Chem. 1991, 56, 3271-3278.-   Pilli et al., J. Org. Chem. 1998, 63, 7811-7819.-   Pulukuri et al., Org. Lett. 2012, 14, 2858-2861.-   Smith, et al., Biochemistry 1995, 34, 415-425.-   Sousa, et al., Tetrahedron 2002, 58, 4643-4654.-   Sun, et al., Biochemistry 1994, 34, 8068-8074.-   S̆venda, et al., Proc. Natl. Acad. Sci. U.S.A. 2011, 108, 6709-6714.-   Tomita, et al., J. Antibiot. 1981a, 34, 1519-1524.-   Tomita, et al., J. Antibiot. 1981b, 34, 1525-1530.-   Wasserman, et al., J. Am. Chem. Soc. 1969, 91, 3674-3675.-   Wasserman, et al., Tetrahedron Lett. 1986a, 27, 4909-4912.-   Wasserman, et al., Tetrahedron Lett. 1986b, 27, 4913-4916.-   Wasserman, et al., Tetrahedron Lett. 1988a, 29, 4973-4976.-   Wasserman, et al., Tetrahedron Lett. 1988b, 29, 4977-4980.-   Yang, et al., J. Am. Chem. Soc. 2009, 133, 12433-12435.

What is claimed is:
 1. A compound of the formula:

wherein: R₁ is amino, hydroxy, or mercapto; alkoxy_((C≤12)),cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),acyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), acylthio_((C≤12)),alkylamino_((C≤12)), cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),alkynylamino_((C≤12)), dialkylamino_((C≤12)),dicycloalkylamino_((C≤12)), dialkenylamino_((C≤12)),dialkynylamino_((C≤12)), amido_((C≤12)), or a substituted version of anyof these groups; or R₁ is a group of the formula:—O-alkanediyl_((C≤8))-alkoxy_((C≤12)),—O-alkanediyl_((C≤8))-alkenyloxy_((C≤12)),—O-alkanediyl_((C≤8))-alkynyloxy_((C≤12)), or a substituted versionthereof; or R₁ is a group of the formula:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein: R_(a) is hydrogen, alkyl_((C≤6)), or substituted alkyl_((C≤6));R₂ and R₃ are independently hydrogen, amino, hydroxy, mercapto;alkyl_((C≤12)), cycloalkyl_((C≤12)), alkenyl_((C≤12)), alkynyl_((C≤12)),alkoxy_((C≤12)), cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)),alkynyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), alkylamino_((C≤12)),cycloalkylamino_((C≤12)), alkenylamino_((C≤12)), alkynylamino_((C≤12)),or a substituted version of any of these groups; R₂ and R₃ are takentogether and are alkoxydiyl_((C≤8)), alkylaminodiyl_((C≤12)),alkylthiodiyl_((C≤12)), or a substituted version of any of these groups;R₄ is hydrogen, amino, halo, hydroxy, mercapto, alkyl_((C≤12)) orsubstituted alkyl_((C≤12)); X₁ and X₂ are each independently hydrogen,hydroxy, or alkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),or a substituted version of any of these groups; and A is a fusedcycloalkanediyl and has the structure:

wherein: Y₁ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₁ is oxo, then the atom to which Y₁is bound is part of a double bond, and provided that when the atom towhich Y₁ is bound is part of a double bond, then Y₁ is oxo; Y₂ ishydrogen, hydroxy, alkyl_((C≤12)), substituted alkyl_((C≤12)),alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or —OX₃, wherein X₃ is ahydroxy protecting group; or a group of the formula:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein:  R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and  R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein:  R_(a) is hydrogen, alkyl_((C≤6)), or substitutedalkyl_((C≤6)); and n₁ is 0, 1, 2, 3, 4, 5, or 6; or A is a fusedarenediyl and has the structure:

wherein: Y₃ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₄ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; and n₂ is 0, 1, 2,or 3; or A is a fused arenediyl with a fused heterocycloalkanediyl andhas the structure:

wherein: Y₅ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₆ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; Y₇ is hydrogen,alkyl_((C≤12)), or substituted alkyl_((C≤12)); n₃ is 0 or 1; and x is 1,2, 3, or 4; or A is a fused heteroarenediyl and has the structure:

wherein: Z₁, Z₂, and Z₃ are each independently selected from CR₅R₅′,NR₅″, O, or S; R₅ and R₅′ are each independently hydrogen, amino,hydroxy, halo, cyano, nitro, sulfato, sulfamido;  alkyl_((C≤6)),alkoxy_((C≤6)), alkylamino_((C≤6)), dialkylamino_((C≤12)),amido_((C≤6)), or a substituted version of any of these groups; and R₅″is hydrogen, alkyl_((C≤12)), or substituted alkyl_((C≤12)); providedthat at least one of Z₁, Z₂, or Z₃ is NR₅″, O, or S; n₄ is 1, 2, 3, or4; or A is a fused arenediyl with a fused cycloalkanediyl and has thestructure:

wherein: Y₈ and Y₉ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₀ is hydrogen, oxo, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; n₅ is 0 or 1; and y is 0, 1, 2, 3, 4, 5, 6, 7, or 8; A is a fusedarenediyl and has the structure:

wherein: Y₁₁ and Y₁₂ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₃ is hydrogen, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; and n₆ is 0, 1, 2, 3, or 4; provided that R₁ is not hydroxy andeither R₂ or R₃ is methoxy when A is a fused cycloalkanediyl of theformula:

or a pharmaceutically acceptable salt thereof.
 2. The compound of claim1 further defined as:

wherein: R₁ is amino, hydroxy, or mercapto; alkoxy_((C≤12)),cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),acyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), acylthio_((C≤12)),alkylamino_((C≤12)), cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),alkynylamino_((C≤12)), dialkylamino_((C≤12)),dicycloalkylamino_((C≤12)), dialkenylamino_((C≤12)),dialkynylamino_((C≤12)), amido_((C≤12)), or a substituted version of anyof these groups; or R₁ is a group of the formula:—O-alkanediyl_((C≤8))-alkoxy_((C≤12)),—O-alkanediyl_((C≤8))-alkenyloxy_((C≤12)),—O-alkanediyl_((C≤8))-alkynyloxy_((C≤12)), or a substituted versionthereof; or R₁ is a group of the formula:

wherein: R₆, R₇, R₈, and R₉ are each independently hydrogen, hydroxy,alkyl_((C≤8)), alkoxy_((C≤8)), substituted alkyl_((C≤8)), or substitutedalkoxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; R₂and R₃ are independently hydrogen, amino, hydroxy, mercapto;alkyl_((C≤12)), cycloalkyl_((C≤12)), alkenyl_((C≤12)), alkynyl_((C≤12)),alkoxy_((C≤12)), cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)),alkynyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), alkylamino_((C≤12)),cycloalkylamino_((C≤12)), alkenylamino_((C≤12)), alkynylamino_((C≤12)),or a substituted version of any of these groups; R₂ and R₃ are takentogether and are alkoxydiyl_((C≤8)), alkylaminodiyl_((C≤12)),alkylthiodiyl_((C≤12)), or a substituted version of any of these groups;R₄ is hydrogen, amino, halo, hydroxy, mercapto, alkyl_((C≤12)) orsubstituted alkyl_((C≤12)); X₁ and X₂ are each independently hydrogen,hydroxy, or alkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),or a substituted version of any of these groups; and A is a fusedcycloalkanediyl and has the structure:

wherein: Y₁ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₁ is oxo, then the atom to which Y₁is bound is part of a double bond, and provided that when the atom towhich Y₁ is bound is part of a double bond, then Y₁ is oxo; Y₂ ishydrogen, hydroxy, alkyl_((C≤12)), substituted alkyl_((C≤12)),alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or —OX₃, wherein X₃ is ahydroxy protecting group; and n₁ is 0, 1, 2, 3, 4, 5, or 6; or A is afused arenediyl and has the structure:

wherein: Y₃ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₄ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; and n₂ is 0, 1, 2,or 3; or A is a fused arenediyl with a fused heterocycloalkanediyl andhas the structure:

wherein: Y₅ is hydrogen, oxo, alkoxy_((C≤12)), or substitutedalkoxy_((C≤12)), provided that when Y₃ is oxo, then the atom to which Y₃is bound is part of a double bond, and provided that when the atom towhich Y₃ is bound is part of a double bond, then Y₃ is oxo; Y₆ ishydrogen, hydroxy, amino, mercapto, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), alkylthio_((C≤12)), substituted alkylthio_((C≤12)),alkylamino_((C≤12)), substituted alkylamino_((C≤12)), —OX₃, wherein X₃is a hydroxy protecting group, —SX₄, wherein X₄ is a thio protectinggroup, or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amineprotecting group and the other is a hydrogen or X₅ and X₆ are takentogether and are a divalent amine protecting group; Y₇ is hydrogen,alkyl_((C≤12)), or substituted alkyl_((C≤12)); n₃ is 0 or 1; and x is 1,2, 3, or 4; or A is a fused heteroarenediyl and has the structure:

wherein: Z₁, Z₂, and Z₃ are each independently selected from CR₅R₅′,NR₅″, O, or S; R₅ and R₅′ are each independently hydrogen, amino,hydroxy, halo, cyano, nitro, sulfato, sulfamido;  alkyl_((C≤6)),alkoxy_((C≤6)), alkylamino_((C≤6)), dialkylamino_((C≤12)),amido_((C≤6)), or a substituted version of any of these groups; and R₅″is hydrogen, alkyl_((C≤12)), or substituted alkyl_((C≤12)); providedthat at least one of Z₁, Z₂, or Z₃ is NR₅″, O, or S; n₄ is 1, 2, 3, or4; or A is a fused arenediyl with a fused cycloalkanediyl and has thestructure:

wherein: Y₈ and Y₉ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₀ is hydrogen, oxo, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; n₅ is 0 or 1; and y is 0, 1, 2, 3, 4, 5, 6, 7, or 8; A is a fusedarenediyl and has the structure:

wherein: Y₁₁ and Y₁₂ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group; Y₁₃ is hydrogen, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃, wherein X₃ is a hydroxyprotecting group, —SX₄, wherein X₄ is a thio protecting group, or—NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protecting groupand the other is a hydrogen or X₅ and X₆ are taken together and are adivalent amine protecting group, provided that when Y₃ is oxo, then theatom to which Y₃ is bound is part of a double bond, and provided thatwhen the atom to which Y₃ is bound is part of a double bond, then Y₃ isoxo; and n₆ is 0, 1, 2, 3, or 4; provided that R₁ is not hydroxy andeither R₂ or R₃ is methoxy when A is a fused cycloalkanediyl of theformula:

or a pharmaceutically acceptable salt thereof.
 3. The compound of eitherclaim 1 or claim 2, wherein the formula is further defined as Ia.
 4. Thecompound of either claim 1 or claim 2, wherein the formula is furtherdefined as Ib.
 5. The compound of either claim 1 or claim 2, wherein theformula is further defined as Ic.
 6. The compound of either claim 1 orclaim 2, wherein the formula is further defined as Id.
 7. The compoundof either claim 1 or claim 2, wherein the formula is further defined asIe.
 8. The compound of either claim 1 or claim 2, wherein the formula isfurther defined as If.
 9. The compound according to any one of claim 1or 3-8, wherein R₁ is:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein: R_(a) is hydrogen, alkyl_((C≤6)), or substituted alkyl_((C≤6)).10. The compound of claim 9, wherein R₁ is:

wherein: R₆, R₇, R₈, and R₉ are each independently hydrogen, hydroxy,alkyl_((C≤8)), alkoxy_((C≤8)), substituted alkyl_((C≤8)), or substitutedalkoxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O-alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group.11. The compound of claim 10, wherein the thiol reactive group of R₁₂ isa maleimide.
 12. The compound according to any one of claims 1-8,wherein R₁ is a group of the formula:—O-alkanediyl_((C≤8))-alkoxy_((C≤12)),—O-alkanediyl_((C≤8))-alkenyloxy_((C≤12)),—O-alkanediyl_((C≤8))-alkynyloxy_((C≤12)), or a substituted versionthereof.
 13. The compound of claim 12, wherein the alkanediyl_((C≤8)) ofR₁ is —CH₂—.
 14. The compound according to any one of claims 1-8,wherein R₁ is alkoxy_((C≤12)) or substituted alkoxy_((C≤12)).
 15. Thecompound of claim 14, wherein R₁ is alkoxy_((C≤12)).
 16. The compound ofclaim 14, wherein R₁ is substituted alkoxy_((C≤12)).
 17. The compound ofclaim 15, wherein R₁ is —O(CH₂)₆OH or —OCH₂CH₂SH.
 18. The compoundaccording to any one of claims 1-8, wherein R₁ is alkynyloxy_((C≤12)) orsubstituted alkynyloxy_((C≤12)).
 19. The compound of claim 18, whereinR₁ is alkynyloxy_((C≤12)).
 20. The compound of claim 19, wherein R₁ is—CH₂CCH.
 21. The compound according to any one of claims 1-7, wherein R₁is alkylthio_((C≤12)) or substituted alkylthio_((C≤12)).
 22. Thecompound of claim 21, wherein R₁ is alkylthio_((C≤12)).
 23. The compoundof claim 22, wherein R₁ is —SCH₂CH₃.
 24. The compound according to anyone of claims 1-23, wherein R₂ is hydrogen.
 25. The compound accordingto any one of claims 1-23, wherein R₂ is alkyl_((C≤12)) or substitutedalkyl_((C≤12)).
 26. The compound of claim 25, wherein R₂ isalkyl_((C≤12)).
 27. The compound of claim 26, wherein R₂ is methyl. 28.The compound according to any one of claims 1-23, wherein R₂ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)).
 29. The compound ofclaim 28, wherein R₂ is alkoxy_((C≤12)).
 30. The compound of claim 29,wherein R₂ is methoxy.
 31. The compound according to any one of claims1-23, wherein R₂ is alkylthio_((C≤12)) or substitutedalkylthio_((C≤12)).
 32. The compound of claim 31, wherein R₂ isalkylthio_((C≤12)).
 33. The compound of claim 32, wherein R₂ is —SCH₃.34. The compound according to any one of claims 1-23, wherein R₂ and R₃are taken together and is alkoxydiyl_((C≤12)) or substitutedalkoxydiyl_((C≤12)).
 35. The compound of claim 34, wherein R₂ and R₃ aretaken together and are alkoxydiyl_((C≤12)).
 36. The compound of claim35, wherein R₂ and R₃ are —OCH₂CH₂O—, —OCH₂CH₂CH₂O—, or—OCH₂C(CH₃)₂CH₂O—.
 37. The compound of claim 34, wherein R₂ and R₃ aretaken together and are alkoxydiyl_((C≤12)).
 38. The compound of claim35, wherein R₂ and R₃ are —OCH₂CH(CH₂OH)CH₂O—, —OCH₂CH(CH₂SH)CH₂O—, or—OCH₂CH(CH₂NHAc)CH₂O—.
 39. The compound according to any one of claims1-23, wherein R₂ and R₃ are taken together and is alkylthiodiyl_((C≤12))or substituted alkylthiodiyl_((C≤12)).
 40. The compound of claim 39,wherein R₂ and R₃ are taken together and are alkylthiodiyl_((C≤12)). 41.The compound of claim 40, wherein R₂ and R₃ are —SCH₂CH₂CH₂S— or—SCH₂C(CH₃)₂CH₂S—.
 42. The compound according to any one of claims 1-33,wherein R₃ is hydrogen.
 43. The compound according to any one of claims1-33, wherein R₃ is alkyl_((C≤12)) or substituted alkyl_((C≤12)). 44.The compound of claim 43, wherein R₃ is alkyl_((C≤12)).
 45. The compoundof claim 44, wherein R₃ is methyl.
 46. The compound according to any oneof claims 1-33, wherein R₃ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)).
 47. The compound of claim 46, wherein R₃ isalkoxy_((C≤12)).
 48. The compound of claim 47, wherein R₃ is methoxy.49. The compound according to any one of claims 1-33, wherein R₃ isalkylthio_((C≤12)) or substituted alkylthio_((C≤12)).
 50. The compoundof claim 49, wherein R₃ is alkylthio_((C≤12)).
 51. The compound of claim50, wherein R₃ is —SCH₃.
 52. The compound according to any one of claims1-51, wherein R₄ is halo.
 53. The compound of claim 52, wherein R₄ isfluoro, chloro, or bromo.
 54. The compound of claim 53, wherein R₄ isfluoro.
 55. The compound according to any one of claims 1-51, wherein R₄is alkyl_((C≤12)) or substituted alkyl_((C≤12)).
 56. The compound ofclaim 55, wherein R₄ is alkyl_((C≤12)).
 57. The compound of claim 56,wherein R₄ is methyl.
 58. The compound of claim 55, wherein R₄ issubstituted alkyl_((C≤12)).
 59. The compound of claim 58, wherein R₄ istrifluoromethyl.
 60. The compound according to any one of claims 1-59,wherein X₁ is hydrogen.
 61. The compound according to any one of claims1-59, wherein X₁ is hydroxy.
 62. The compound according to any one ofclaims 1-59, wherein X₁ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)).
 63. The compound of claim 62, wherein X₁ isalkoxy_((C≤12)).
 64. The compound of claim 63, wherein X₁ is methoxy.65. The compound of claim 62, wherein X₁ is substituted alkoxy_((C≤12)).66. The compound of claim 65, wherein X₁ is —O(CH₂)₃NH₂,—O(CH₂)₂C(O)NH₂, or —O(CH₂)₃SH.
 67. The compound according to any one ofclaims 1-59, wherein X₁ is alkenyloxy_((C≤12)).
 68. The compound ofclaim 67, wherein X₁ is —OCH₂CHCH₂.
 69. The compound according to anyone of claims 1-59, wherein X₁ is alkynyloxy_((C≤12)).
 70. The compoundof claim 69, wherein X₁ is —OCH₂CCH.
 71. The compound according to anyone of claims 1-64, wherein X₂ is hydrogen.
 72. The compound accordingto any one of claims 1-64, wherein X₂ is hydroxy.
 73. The compoundaccording to any one of claims 1-64, wherein X₂ is alkoxy_((C≤12)) orsubstituted alkoxy_((C≤12)).
 74. The compound of claim 73, wherein X₂ isalkoxy_((C≤12)).
 75. The compound of claim 74, wherein X₂ is methoxy.76. The compound of claim 1-3 or 14-75, wherein Y₁ is oxo.
 77. Thecompound of claim 1-3 or 14-76, wherein Y₂ is hydrogen.
 78. The compoundaccording to any one of claim 1-3, or 14-76, wherein Y₂ is hydroxy. 79.The compound according to any one of claim 1-3 or 14-76, wherein Y₂ is:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyl_((C≤12)), substituted alkyl_((C≤12)), substitutedalkoxy_((C≤12)), substituted acyl_((C≤12)), or a group of the formula:

wherein: R₁₁ is hydrogen, alkyl_((C≤8)), or substituted alkyl_((C≤8));and R₁₂ is hydrogen, hydroxy, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), —O— alkanediyl_((C≤12))-a thiol reactive group, or asubstituted version of —O-alkanediyl_((C≤12))-a thiol reactive group; orR₇ and R₁₀ are taken together to form a heterocyclic compound of theformula:

wherein: R_(a) is hydrogen, alkyl_((C≤6)), or substituted alkyl_((C≤6)).80. The compound of claim 79, wherein Y₂ is:

wherein: R₆, R₆′, R₇, R₈, R₈′, R₉, and R₉′ are each independentlyhydrogen, hydroxy, alkyl_((C≤8)), alkoxy_((C≤8)), acyloxy_((C≤8)),substituted alkyl_((C≤8)), substituted alkoxy_((C≤8)), or substitutedacyloxy_((C≤8)); and R₁₀ is hydrogen, hydroxy, alkyl_((C≤12)),alkoxy_((C≤12)), acyloxy_((C≤12)), acyl_((C≤12)), substitutedalkyl_((C≤12)), substituted alkoxy_((C≤12)), substitutedacyloxy_((C≤12)), or substituted acyl_((C≤12)).
 81. The compound ofeither claim 79 or 80, wherein Y₂ is:


82. The compound according to any one of claim 1-3 or 14-81, wherein n₁is 0, 1, 2, or
 3. 83. The compound of claim 82, wherein n₁ is 0, 1, or2.
 84. The compound according to any one of claim 1, 4, or 14-75,wherein Y₃ is hydroxy.
 85. The compound according to any one of claim 1,4, or 14-75, wherein Y₃ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)).
 86. The compound of claim 85, wherein Y₃ isalkoxy_((C≤12)).
 87. The compound of claim 86, wherein Y₃ is methoxy.88. The compound of claim 85, wherein Y₃ is substituted alkoxy_((C≤12)).89. The compound of claim 88, wherein Y₃ is methoxymethoxy.
 90. Thecompound according to any one of claim 1, 4, 14-75, or 84-89, wherein Y₄is hydrogen.
 91. The compound according to any one of claim 1, 4, 14-75,or 84-89, wherein Y₄ is hydroxy.
 92. The compound according to any oneof claim 1, 4, 14-75, or 84-89, wherein Y₄ is alkoxy_((C≤12)) orsubstituted alkoxy_((C≤12)).
 93. The compound of claim 92, wherein Y₄ isalkoxy_((C≤12)).
 94. The compound of claim 93, wherein Y₄ is methoxy.95. The compound according to any one of claim 1, 4, 14-75, or 84-89,wherein Y₄ is alkylamino_((C≤12)) or substituted alkylamino_((C≤12)).96. The compound of claim 95, wherein Y₄ is alkylamino_((C≤12)).
 97. Thecompound of claim 96, wherein Y₄ is methylamino.
 98. The compoundaccording to any one of claim 1, 4, 14-75, or 84-97, wherein n₂ is 1, 2,or
 3. 99. The compound according to any one of claim 1, 5, or 14-75,wherein Y₅ is hydroxy.
 100. The compound according to any one of claim1, 5, or 14-75, wherein Y₅ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)).
 101. The compound of claim 100, wherein Y₅ isalkoxy_((C≤12)).
 102. The compound of claim 101, wherein Y₅ is methoxy.103. The compound according to any one of claim 1, 5, 14-75, or 99-102,wherein Y₆ is hydrogen.
 104. The compound according to any one of claim1, 5, 14-75, or 99-102, wherein Y₆ is hydroxy.
 105. The compoundaccording to any one of claim 1, 5, 14-75, or 99-102, wherein Y₆ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)).
 106. The compound ofclaim 105, wherein Y₆ is alkoxy_((C≤12)).
 107. The compound of claim106, wherein Y₆ is methoxy.
 108. The compound according to any one ofclaim 1, 5, 14-75, or 99-107, wherein Y₇ is hydrogen.
 109. The compoundaccording to any one of claim 1, 5, 14-75, or 99-107, wherein Y₇ isalkyl_((C≤6)) or substituted alkyl_((C≤6)).
 110. The compound accordingto any one of claim 1, 5, 14-75, or 99-109, wherein x is 2 or
 3. 111.The compound of claim 110, wherein x is
 2. 112. The compound of claim110, wherein x is
 3. 113. The compound according to any one of claim 1,5, 14-75, or 99-109, wherein n₃ is
 0. 114. The compound according to anyone of claim 1, 5, 14-75, or 99-109, wherein n₃ is
 1. 115. The compoundaccording to any one of claim 1, 6, or 14-75, wherein Z₁ is S.
 116. Thecompound according to any one of claim 1, 6, or 14-75, wherein Z₁ is N.117. The compound according to any one of claim 1, 6, or 14-75, whereinZ₁ is O.
 118. The compound according to any one of claim 1, 6, 14-75, or115-117, wherein Z₂ is S.
 119. The compound according to any one ofclaim 1, 6, 14-75, or 115-117, wherein Z₂ is N.
 120. The compoundaccording to any one of claim 1, 6, 14-75, or 115-117, wherein Z₂ is O.121. The compound according to any one of claim 1, 6, 14-75, or 115-117,wherein Z₂ is CR₅″.
 122. The compound of claim 121, wherein R₅″ ishydrogen, hydroxy, halo, alkyl_((C≤12)), substituted alkyl_((C≤12)),alkoxy_((C≤12)), or substituted alkoxy_((C≤12)).
 123. The compound ofclaim 122, wherein R₅″ is hydrogen.
 124. The compound of claim 122,wherein R₅″ is alkoxy_((C≤12)) or substituted alkoxy_((C≤12)).
 125. Thecompound of claim 124, wherein R₅″ is methoxy.
 126. The compound ofclaim 122, wherein R₅″ is alkyl_((C≤12)) or substituted alkyl_((C≤12)).127. The compound of claim 126, wherein R₅″ is methyl.
 128. The compoundaccording to any one of claim 1, 6, 14-75, or 115-128, wherein Z₃ is S.129. The compound according to any one of claim 1, 6, 14-75, or 115-128,wherein Z₃ is N.
 130. The compound according to any one of claim 1, 6,14-75, or 115-128, wherein Z₃ is O.
 131. The compound according to anyone of claim 1, 6, 14-75, or 115-128, wherein Z₃ is CR₅″.
 132. Thecompound of claim 131, wherein R₅″ is hydrogen, hydroxy, halo,alkyl_((C≤12)), substituted alkyl_((C≤12)), alkoxy_((C≤12)), orsubstituted alkoxy_((C≤12)).
 133. The compound of claim 131, wherein R₅″is hydrogen.
 134. The compound of claim 132, wherein R₅″ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)).
 135. The compound ofclaim 134, wherein R₅″ is methoxy.
 136. The compound of claim 132,wherein R₅″ is alkyl_((C≤12)) or substituted alkyl_((C≤12)).
 137. Thecompound of claim 136, wherein R₅″ is methyl.
 138. The compoundaccording to any one of claim 1, 6, 14-75, or 115-137, wherein n₄ is 1,2, or
 3. 139. The compound according to any one of claims 1, 7, 14-75,wherein Y₈ is hydrogen.
 140. The compound according to any one of claims1, 7, 14-75, wherein Y₈ is hydroxy.
 141. The compound according to anyone of claims 1, 7, 14-75, wherein Y₈ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)).
 142. The compound of claim 141, wherein Y₈ isalkoxy_((C≤12)).
 143. The compound of claim 142, wherein Y₈ is methoxy.144. The compound according to any one of claim 1, 7, 14-75, or 139-143,wherein Y₉ is hydrogen.
 145. The compound according to any one of claim1, 7, 14-75, or 139-143, wherein Y₉ is hydroxy.
 146. The compoundaccording to any one of claim 1, 7, 14-75, or 139-143, wherein Y₉ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)).
 147. The compound ofclaim 146, wherein Y₉ is alkoxy_((C≤12)).
 148. The compound of claim147, wherein Y₉ is methoxy.
 149. The compound according to any one ofclaim 1, 7, 14-75, or 139-148, wherein Y₁₀ is hydrogen.
 150. Thecompound according to any one of claim 1, 7, 14-75, or 139-148, whereinY₁₀ is hydroxy.
 151. The compound according to any one of claim 1, 7,14-75, or 139-148, wherein Y₁₀ is oxo.
 152. The compound according toany one of claim 1, 7, 14-75, or 139-148, wherein Y₁₀ is alkoxy_((C≤12))or substituted alkoxy_((C≤12)).
 153. The compound of claim 152, whereinY₁₀ is alkoxy_((C≤12)).
 154. The compound of claim 153, wherein Y₁₀ ismethoxy.
 155. The compound according to any one of claim 1, 8, 14-75, or139-148, wherein Y₁₀ is alkylamino_((C≤12)) or substitutedalkylamino_((C≤12)).
 156. The compound of claim 146, wherein Y₁₀ isalkylamino_((C≤12)).
 157. The compound of claim 147, wherein Y₁₀ ismethylamino.
 158. The compound according to any one of claim 1, 7,14-75, or 139-154, wherein n₅ is
 1. 159. The compound according to anyone of claim 1, 7, 14-75, or 139-158, wherein y is 1, 2, 3, 4, 5, or 6.160. The compound according to any one of claims 1, 8, 14-75, whereinY₁₁ is hydrogen.
 161. The compound according to any one of claims 1, 8,14-75, wherein Y₁₁ is hydroxy.
 162. The compound according to any one ofclaims 1, 8, 14-75, wherein Y₁₁ is alkoxy_((C≤12)) or substitutedalkoxy_((C≤12)).
 163. The compound of claim 141, wherein Y₁₁ isalkoxy_((C≤12)).
 164. The compound of claim 142, wherein Y₁₁ is methoxy.165. The compound according to any one of claim 1, 8, 14-75, or 160-164,wherein Y₁₂ is hydrogen.
 166. The compound according to any one of claim1, 8, 14-75, or 160-164, wherein Y₁₂ is hydroxy.
 167. The compoundaccording to any one of claim 1, 8, 14-75, or 160-164, wherein Y₁₂ isalkoxy_((C≤12)) or substituted alkoxy_((C≤12)).
 168. The compound ofclaim 146, wherein Y₁₂ is alkoxy_((C≤12)).
 169. The compound of claim147, wherein Y₁₂ is methoxy.
 170. The compound according to any one ofclaim 1, 8, 14-75, or 160-169, wherein Y₁₃ is hydrogen.
 171. Thecompound according to any one of claim 1, 8, 14-75, or 160-169, whereinY₁₃ is hydroxy.
 172. The compound according to any one of claim 1, 8,14-75, or 160-169, wherein Y₁₃ is oxo.
 173. The compound according toany one of claim 1, 8, 14-75, or 160-169, wherein Y₁₃ is alkoxy_((C≤12))or substituted alkoxy_((C≤12)).
 174. The compound of claim 152, whereinY₁₃ is alkoxy_((C≤12)).
 175. The compound of claim 153, wherein Y₁₃ ismethoxy.
 176. The compound according to any one of claim 1, 8, 14-75, or160-169, wherein Y₁₃ is alkylamino_((C≤12)) or substitutedalkylamino_((C≤12)).
 177. The compound of claim 146, wherein Y₁₃ isalkylamino_((C≤12)).
 178. The compound of claim 147, wherein Y₁₃ ismethylamino.
 179. The compound according to any one of claim 1, 8,14-75, or 160-154, wherein n₆ is 1, 2, or
 3. 180. The compound of claim179, wherein n₆ is 2 or
 3. 181. The compound according to any one ofclaims 1-180, wherein the compound is further defined as:

or a pharmaceutically acceptable salt thereof.
 182. A pharmaceuticalcomposition comprising a compound according to any one of claims 1-181and a pharmaceutically acceptable carrier.
 183. The pharmaceuticalcomposition of claim 182, wherein the composition is formulated foradministration: orally, intraadiposally, intraarterially,intraarticularly, intracranially, intradermally, intralesionally,intramuscularly, intranasally, intraocularly, intrapericardially,intraperitoneally, intrapleurally, intraprostatically, intrarectally,intrathecally, intratracheally, intratumorally, intraumbilically,intravaginally, intravenously, intravesicularlly, intravitreally,liposomally, locally, mucosally, parenterally, rectally,subconjunctival, subcutaneously, sublingually, topically, transbuccally,transdermally, vaginally, in crémes, in lipid compositions, via acatheter, via a lavage, via continuous infusion, via infusion, viainhalation, via injection, via local delivery, or via localizedperfusion.
 184. A method of treating a disease or disorder in a patientin need thereof comprising administering to the patient apharmaceutically effective amount of a compound or composition accordingto any one of claims 1-183.
 185. The method of claim 184, wherein thedisease or disorder is cancer.
 186. The method of claim 185, wherein thecancer is a carcinoma, sarcoma, lymphoma, leukemia, melanoma,mesothelioma, multiple myeloma, or seminoma.
 187. The method of claim185, wherein the cancer is of the bladder, blood, bone, brain, breast,central nervous system, cervix, colon, endometrium, esophagus, gallbladder, gastrointestinal tract, genitalia, genitourinary tract, head,kidney, larynx, liver, lung, muscle tissue, neck, oral or nasal mucosa,ovary, pancreas, prostate, skin, spleen, small intestine, largeintestine, stomach, testicle, or thyroid.
 188. The method according toany one of claims 184-187 further comprising a second therapeutic agentor modality.
 189. The method according to any one of claims 184-188,wherein the compound is administered once.
 190. The method according toany one of claims 184-188, wherein the compound is administered two ormore times.
 191. A method of preparing a compound comprising reacting acompound of the formula:

wherein: R₁ is amino, hydroxy, or mercapto; or —OX₃, wherein X₃ is ahydroxy protecting group, —SX₄, wherein X₄ is a thio protecting group,or —NX₅X₆, wherein either X₅ or X₆ is a monovalent amine protectinggroup and the other is a hydrogen or X₅ and X₆ are taken together andare a divalent amine protecting group; R₂ and R₃ are independentlyselected from hydrogen, amino, hydroxy, mercapto; alkyl_((C≤12)),cycloalkyl_((C≤12)), alkenyl_((C≤12)), alkynyl_((C≤12)),alkoxy_((C≤12)), cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)),alkynyloxy_((C≤12)), alkylthio_((C≤12)), cycloalkylthio_((C≤12)),alkenylthio_((C≤12)), alkynylthio_((C≤12)), alkylamino_((C≤12)),cycloalkylamino_((C≤12)), alkenylamino_((C≤12)), alkynylamino_((C≤12)),or a substituted version of any of these groups, or —OX₇, wherein X₇ isa hydroxy protecting group, —SX₈, wherein X₈ is a thio protecting group,or —NX₉X₁₀, wherein either X₉ or X₁₀ is a monovalent amine protectinggroup and the other is a hydrogen or X₉ and X₁₀ are taken together andare a divalent amine protecting group; R₂ and R₃ are taken together andare alkoxydiyl_((C≤8)), alkylaminodiyl_((C≤12)), alkylthiodiyl_((C≤12)),or a substituted version of any of these groups; R₄ is hydrogen, amino,halo, hydroxy, mercapto, alkyl_((C≤12)) or substituted alkyl_((C≤12)),or —OX₁₁, wherein X₁₁ is a hydroxy protecting group, —SX₁₂, wherein X₁₂is a thio protecting group, or —NX₁₃X₁₄, wherein either X₁₃ or X₁₄ is amonovalent amine protecting group and the other is a hydrogen or X₁₃ andX₁₄ are taken together and are a divalent amine protecting group; R₅ ishydrogen or a hydroxy protecting group; R₆ is hydrogen oralkylidene_((C≤12)), alkyl_((C≤12)), cycloalkyl_((C≤12)),alkenyl_((C≤12)), alkynyl_((C≤12)), alkoxy_((C≤12)),cycloalkoxy_((C≤12)), alkenyloxy_((C≤12)), alkynyloxy_((C≤12)),alkylamino_((C≤12)), dialkylamino_((C≤12)), or a substituted version ofany of these groups; X₁ and X₂ are each independently hydrogen, hydroxy,alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or —OX₁₅, wherein X₁₅ is ahydroxy protecting group; or X₁₅ and R₅ are taken together and are adivalent diol protecting group; and A is a fused cycloalkanediyl and hasthe structure:

wherein: Y₁ is hydrogen, oxo, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), or —OX₁₆, wherein X₁₆ is a hydroxy protecting group;provided that when Y₁ is oxo, then the atom to which Y₁ is bound is partof a double bond, and provided that when the atom to which Y₁ is boundis part of a double bond, then Y₁ is oxo; Y₂ is hydrogen, hydroxy,alkoxy_((C≤12)), substituted alkoxy_((C≤12)), or —OX₁₇, wherein X₁₇ is ahydroxy protecting group; and n₁ is 0, 1, 2, 3, 4, 5, or 6; or A is afused arenediyl and has the structure:

wherein: Y₃ is hydrogen, oxo, alkoxy_((C≤12)), substitutedalkoxy_((C≤12)), or —OX₁₈, wherein X₁₈ is a hydroxy protecting group;provided that when Y₃ is oxo, then the atom to which Y₃ is bound is partof a double bond, and provided that when the atom to which Y₃ is boundis part of a double bond, then Y₃ is oxo; Y₄ is hydrogen, hydroxy,amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₁₉, wherein X₁₉ is a hydroxyprotecting group, —SX₂₀, wherein X₂₀ is a thio protecting group, or—NX₂₁X₂₂, wherein either X₂₁ or X₂₂ is a monovalent amine protectinggroup and the other is a hydrogen or X₂₁ and X₂₂ are taken together andare a divalent amine protecting group; and n₂ is 0, 1, 2, or 3; or A isa fused arenediyl with a fused heterocycloalkanediyl and has thestructure:

wherein: Y₅ is hydrogen, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),or —OX₂₃, wherein X₂₃ is a hydroxy protecting group; Y₆ is hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₂₄, wherein X₂₄ is a hydroxyprotecting group, —SX₂₅, wherein X₂₅ is a thio protecting group, or—NX₂₆X₂₇, wherein either X₂₆ or X₂₇ is a monovalent amine protectinggroup and the other is a hydrogen or X₂₆ and X₂₇ are taken together andare a divalent amine protecting group; Y₇ is hydrogen, alkyl_((C≤12)),or substituted alkyl_((C≤12)); n₃ is 0 or 1; and x is 1, 2, 3, or 4; orA is a fused heteroarenediyl and has the structure:

wherein: Z₁, Z₂, and Z₃ are each independently selected from CR₆R₆′,NR₇, O, or S; R₆ and R₆′ are each independently hydrogen, amino,hydroxy, halo, cyano, nitro, sulfato, sulfamido;  alkyl_((C≤6)),alkoxy_((C≤6)), alkylamino_((C≤6)), dialkylamino_((C≤12)),amido_((C≤6)), or a substituted version of any of these groups; and R₇is hydrogen, alkyl_((C≤12)), or substituted alkyl_((C≤12)); providedthat at least one of Z₁, Z₂, or Z₃ is NR₆, O, or S; n₄ is 1, 2, 3, or 4;or A is a fused arenediyl with a fused cycloalkanediyl and has thestructure:

wherein: Y₈ and Y₉ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₂₈, wherein X₂₈ is a hydroxyprotecting group, —SX₂₉, wherein X₂₉ is a thio protecting group, or—NX₃₀X₃₁, wherein either X₃₀ or X₃₁ is a monovalent amine protectinggroup and the other is a hydrogen or X₃₀ and X₃₁ are taken together andare a divalent amine protecting group; Y₁₀ is hydrogen, oxo, hydroxy,amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃₂, wherein X₃₂ is a hydroxyprotecting group, —SX₃₃, wherein X₃₃ is a thio protecting group, or—NX₃₄X₃₅, wherein either X₃₄ or X₃₅ is a monovalent amine protectinggroup and the other is a hydrogen or X₃₄ and X₃₅ are taken together andare a divalent amine protecting group, provided that when Y₁₀ is oxo,then the atom to which Y₁₀ is bound is part of a double bond, andprovided that when the atom to which Y₁₀ is bound is part of a doublebond, then Y₁₀ is oxo; n₅ is 0 or 1; and y is 0, 1, 2, 3, 4, 5, 6, 7, or8; A is a fused arenediyl and has the structure:

wherein: Y₁₁ and Y₁₂ are each independently selected from hydrogen,hydroxy, amino, mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₃₆, wherein X₃₆ is a hydroxyprotecting group, —SX₃₇, wherein X₃₇ is a thio protecting group, or—NX₃₈X₃₉, wherein either X₃₈ or X₃₉ is a monovalent amine protectinggroup and the other is a hydrogen or X₃₈ and X₃₉ are taken together andare a divalent amine protecting group; Y₁₃ is hydrogen, hydroxy, amino,mercapto, alkoxy_((C≤12)), substituted alkoxy_((C≤12)),alkylthio_((C≤12)), substituted alkylthio_((C≤12)), alkylamino_((C≤12)),substituted alkylamino_((C≤12)), —OX₄₀, wherein X₄₀ is a hydroxyprotecting group, —SX₄₁, wherein X₄₁ is a thio protecting group, or—NX₄₂X₄₃, wherein either X₄₂ or X₄₃ is a monovalent amine protectinggroup and the other is a hydrogen or X₄₂ and X₄₃ are taken together andare a divalent amine protecting group; and n₆ is 0, 1, 2, 3, or 4; witha Lewis acid under conditions sufficient to produce a compound of theformula:

wherein: R₁, R₂, R₃, R₄, R₅, X₁, and X₂ are as defined above; A is afused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

or wherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyland has the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; or a salt thereof.192. The method of claim 191, wherein the Lewis acid is a transitionmetal or a boron complex.
 193. The method of claim 191 or claim 192,wherein the Lewis acid is a boron complex.
 194. The method according toany one of claim 191-193, wherein the Lewis acid is boron trifluorideetherate.
 195. The method of claim 191 or claim 192, wherein the Lewisacid is a transition metal complex.
 196. The method according to any oneof claim 191, 192, or 195, wherein the Lewis acid is SnCl₄.
 197. Themethod according to any one of claims 191-196, wherein the methodfurther comprises a solvent.
 198. The method of claim 197, wherein thesolvent is chloroalkane_((C≤12)).
 199. The method of claim 198, whereinthe solvent is dichloromethane.
 200. The method according to any one ofclaims 191-196, wherein R₆ is alkylidene_((C≤12)) or substitutedalkyldiene_((C≤12)).
 201. The method according to any one of claims191-199, wherein the method further comprises reacting the compound withan epoxidizing agent under conditions to sufficient to produce acompound of the formula:

wherein: R₁, R₂, R₃, R₄, R₅, X₁, and X₂ are as defined above; A is afused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

or wherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyland has the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; or a salt thereof.202. The method of claim 201, wherein the epoxidizing agent is osmiumtetraoxide with tosyl chloride and a base.
 203. The method of claim 202,wherein the osmium tetraoxide is added to the compound and after a timeperiod of about 1 hour to about 24 hours, the tosyl chloride and thebase are added.
 204. The method according to any one of claims 191-203,wherein the method further comprises reacting the compound with anoxidizing agent under conditions sufficient to produce a compound of theformula:

wherein: R₂, R₃, R₄, R₅, X₁, and X₂ are as defined above; R₁ is O, S, orNH; and A is a fused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

or wherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyland has the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; or a salt thereof.205. The method of claim 204, wherein the oxidizing agent istetrapropylammonium perruthenate and N-methylmorpholine N-oxide. 206.The method according to any one of claims 191-205, wherein the methodfurther comprises reacting the compound with a fluoride source undercondition sufficient to produce a compound of the formula:

wherein: R₂, R₃, R₄, X₁, and X₂ are as defined above; R₁ is amino,hydroxy, or mercapto; alkoxy_((C≤12)), cycloalkoxy_((C≤12)),alkenyloxy_((C≤12)), alkynyloxy_((C≤12)), alkylthio_((C≤12)),cycloalkylthio_((C≤12)), alkenylthio_((C≤12)), alkynylthio_((C≤12)),alkylamino_((C≤12)), cycloalkylamino_((C≤12)), alkenylamino_((C≤12)),alkynylamino_((C≤12)), dialkylamino_((C≤12)),dicycloalkylamino_((C≤12)), dialkenylamino_((C≤12)),dialkynylamino_((C≤12)), or a substituted version of any of thesegroups; A is a fused cycloalkanediyl and has the structure:

wherein: Y₁, Y₂, and n₁ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₃, Y₄, and n₂ are as defined above; A is a fused arenediylwith a fused heterocycloalkanediyl and has the structure:

wherein: Y₅, Y₆, Y₇, x, and n₃ are as defined above; A is a fusedheteroarenediyl and has the structure:

wherein: Z₁, Z₂, Z₃, and n₄ are as defined above; A is a fused arenediylwith a fused cycloalkanediyl and has the structure:

wherein: Y₈, Y₉, and n₅ are as defined above; A is a fused arenediyl andhas the structure:

wherein: Y₁₁, Y₁₂, Y₁₃, and n₆ are as defined above; or a salt thereof.207. The method of claim 206, wherein the fluoride source is Et₃N.3HF.208. The method according to any one of claims 191-207, wherein themethod further comprises one or more deprotection steps.
 209. Aconjugate of the formula:(A-L)_(n)-X  (VI) wherein: A is a compound according to any one ofclaims 1-181; L is a covalent bond or a linker; n is 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12; and X is a cell targeting moiety.